JP2013155354A - Gelatin vitrigel and method for producing the same, medical material, cosmetic product and food material each using the gelatin vitrigel, gelatin gel dried body, gelatin vitrigel dried body and method for producing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a gelatin vitrigel which is inexpensive and excellent in thermal stability and a method for producing the same, and further to provide a medical material, a cosmetic product and a food material each using the gelatin vitrigel.SOLUTION: The method for producing a gelatin vitrigel mainly composed of gelatin includes the steps of: (1) causing a gelatin aqueous solution containing insoluble collagen or collagen which can be refibriformed to gelate by cooling, thereby preparing a gelatin gel mainly composed of gelatin; (2) drying the gelatin gel to thereby produce a vitrified gelatin gel dried body; (3) irradiating the gelatin gel dried body with ultraviolet ray; and (4) rehydrating the gelatin gel dried body after the step (3) to thereby produce the gelatin vitrigel.

Description

  The present invention relates to gelatin vitrigel and a method for producing the same, medical materials, cosmetics and food materials using gelatin vitrigel, dried gelatin gel, dried gelatin vitrigel, and a method for producing the same.

  The present inventor injected a collagen sol imparted with a salt concentration and hydrogen ion concentration (pH) optimal for gelation at low temperature into a culture dish, and further kept the temperature at an optimal temperature. After the collagen sol is gelled, it is sufficiently dried at low temperature to gradually remove not only free water but also bound water, vitrification, and rehydration, collagen gel A technology has been established for converting the physical properties of the above into a thin film excellent in strength and transparency with good reproducibility (Patent Document 1).

  And if this inventor is a hydrogel, even if it is a gel of components other than collagen, by rehydrating after vitrification, the gel can be converted into a stable new physical property state. The gel of a new physical property state produced through the above is named vitrigel (Non-patent Document 1), and this name has already been established as a new academic term.

  The present inventor has further developed a technique relating to vitrigel to improve the transparency and production reproducibility of the collagen vitrigel membrane (Patent Document 2), and to form the collagen vitrigel in a filamentous or tubular form instead of a membrane form. A technique for producing the shape (Patent Document 3) has also been established.

JP-A-8-228768 WO2005 / 014774 JP 2007-204881

Takezawa T, et al., Cell Transplant. 13: 463-473, 2004

  Thus, for example, various studies have been made on vitrigel using native collagen such as bovine or pig or atelocollagen, and application to various fields is expected. However, since collagen is relatively expensive, In the case of a collagen vitrigel made from collagen, there is a problem that it is difficult to control costs. Therefore, in order to distribute various products applying Vitrigel to the market, it is required to keep the cost low.

  Therefore, the present inventors paid attention to inexpensive gelatin as a raw material for vitrigel. The fact that gelatin can be used as a raw material is also described in, for example, Patent Document 2, but gelatin dissolves at high temperatures. Therefore, gelatin vitrigel made from gelatin is used at a living body temperature (approximately 37 ° C.). In order to apply to medical materials and cosmetics used, it was considered that further improvement in thermal stability was necessary.

  Moreover, if the thermal stability of gelatin vitrigel can be improved, it is considered that, for example, a completely new food material can be created as gelatin that does not dissolve even when cooked.

  The present invention has been made in view of the above circumstances, and provides a gelatin vitrigel that is inexpensive and excellent in thermal stability, and a method for producing the gelatin vitrigel, and further, a medical material using the gelatin vitrigel. It is an object to provide cosmetics and food materials. Another object of the present invention is to provide a dried gelatin gel, a dried gelatin vitrigel gel, and a method for producing the gelatin vitrigel that can provide such a gelatin vitrigel.

In order to solve the above problems, the gelatin vitrigel production method of the present invention is characterized by the following.
<1> A method for producing gelatin vitrigel mainly composed of gelatin, comprising the following steps:
(1) A step of producing a gelatin gel containing gelatin as a main component by cooling an aqueous gelatin solution containing insoluble collagen or collagen that can be refibrinated.
(2) A step of drying a gelatin gel to produce a vitrified gelatin gel dried body;
(3) a step of irradiating the dried gelatin gel with ultraviolet rays; and (4) a step of rehydrating the dried gelatin gel after the step (3) to prepare a gelatin vitrigel.
<2> In the step (3), the both sides of the dried gelatin gel are irradiated with ultraviolet rays.
<3> The amount of ultraviolet light irradiated on both sides of the dried gelatin gel is 800 mJ / cm ² or more for each side of the dried gelatin gel.
<4> The gelatin aqueous solution in the step (1) is a crude extracted gelatin aqueous solution containing insoluble collagen as an insoluble residue derived from animal skin.
<5> The aqueous gelatin solution in the step (1) is a mixed aqueous solution to which an aqueous solution containing insoluble collagen or collagen capable of refibrosis is added after purified gelatin is dissolved in a solvent.
<6> The solvent for dissolving the purified gelatin is an equilibrium salt solution that maintains a physiological salt concentration and a hydrogen ion concentration (pH).

The gelatin vitrigel of the present invention is characterized by the following.
<7> Manufactured by any one of the methods <1> to <6>.
<8> A gelatin vitrigel produced by the method of <4> above, wherein the raw skin of the animal used as a raw material for the crude extracted gelatin aqueous solution is derived from fish.
<9> In the gelatin vitrigel of <8>, the fish is a tuna.

  The medical material of the present invention includes <10> vitrigel manufactured by the method of <5>.

  The cosmetic product of the present invention includes <11> the gelatin vitrigel of any one of <7> to <9> and a cosmetic component.

  The food material of the present invention includes <12> the gelatin vitrigel of any one of <7> to <9> as a raw material.

The method for producing a dried gelatin gel of the present invention is characterized by the following.
<13> A method for producing a dried gelatin gel comprising gelatin as a main component and capable of producing a gelatin vitrigel, comprising the following steps:
(1) A step of producing a gelatin gel containing gelatin as a main component by cooling an aqueous gelatin solution containing insoluble collagen or collagen that can be refibrinated.
(2) A step of drying the gelatin gel to produce a vitrified gelatin gel dried product; and (3) a step of irradiating the gelatin gel dried product with ultraviolet rays.
<14> The gelatin aqueous solution in the step (1) is a crude extracted gelatin aqueous solution containing insoluble collagen as an insoluble residue derived from animal skin.
<15> The aqueous gelatin solution in the step (1) is a mixed aqueous solution to which an aqueous solution containing insoluble collagen or collagen capable of refibrosis is added after the purified gelatin is dissolved in a solvent. Process for producing a dried gelatin gel.

The dried gelatin gel of the present invention is characterized by the following.
<16> Manufactured by any one of the methods <13> to <15>.
<17> A dried gelatin gel produced by the method of <14> above, wherein the raw skin of the animal used as a raw material for the crude extracted gelatin aqueous solution is derived from fish.
<18> In the dried gelatin gel of <17>, the fish is a tuna.

The method for producing a dried gelatin vitrigel of the present invention is characterized by the following.
<19> A method for producing a dried gelatin vitrigel comprising gelatin as a main component, the following steps:
(1) A step of producing a gelatin gel containing gelatin as a main component by cooling an aqueous gelatin solution containing insoluble collagen or collagen that can be refibrinated.
(2) A step of drying a gelatin gel to produce a vitrified gelatin gel dried body;
(3) A step of irradiating the dried gelatin gel with ultraviolet rays;
(4) A step of rehydrating the gelatin gel dried material obtained through the step (3) to prepare a gelatin vitrigel; and (5) Redrying the gelatin vitrigel to produce a vitrified gelatin vitrigel dried material. The process of carrying out is included.
<20> Further, (6) a step of irradiating the dried gelatin vitrigel with ultraviolet rays is included.
<21> In the step (3), one surface of the dried gelatin gel is irradiated with ultraviolet rays, and in the step (6), the other surface of the dried gelatin vitrigel is irradiated with ultraviolet rays.
<22> The gelatin aqueous solution in the step (1) is a crude extracted gelatin aqueous solution containing insoluble collagen as an insoluble residue derived from animal skin.
<23> The gelatin aqueous solution in the step (1) is a mixed aqueous solution to which purified gelatin is dissolved in a solvent and an aqueous solution containing insoluble collagen or collagen capable of refibrosis is added.

The dried gelatin vitrigel of the present invention is characterized by the following.
<24> Manufactured by any one of the methods <19> to <23>.
<25> A dried gelatin vitrigel produced by the method of <22> above, wherein the raw skin of the animal used as a raw material for the crude extracted gelatin aqueous solution is derived from fish.
<26> In the dried gelatin vitrigel according to <25>, the fish is a tuna.

  The gelatin vitrigel of the present invention is obtained by rehydrating <27> a dried gelatin vitrigel according to any one of <24> to <26> above.

  According to the production method of the present invention, it is possible to produce a gelatin vitrigel that is inexpensive and excellent in thermal stability. In addition, the gelatin vitrigel of the present invention is inexpensive and excellent in heat stability, and therefore can be applied to medical materials, cosmetics, food materials, and the like. Furthermore, the dried gelatin gel and dried gelatin vitrigel of the present invention can be converted into gelatin vitrigel by rehydration, and can be effectively used industrially.

2 is a phase contrast micrograph showing an insoluble residue contained in a gelatin solution used in Example 1. FIG. Bar in the figure indicates 0.2 mm. It is a macrophotograph which shows a mode that the gelatin vitrigel produced in Example 1 was picked up with tweezers. 2 is a macroscopic photograph from above of the gelatin vitrigel prepared in Example 1. FIG. It is a macrophotograph which shows the state 2 hours after moving the gelatin vitrigel produced in Example 1 in a 37 degreeC incubator. In Comparative example 1, it is the phase-contrast microscope picture which showed the state which removed the insoluble residue in the gelatin vitrigel used in Example 1 with the filter. Bar in the figure indicates 0.2 mm. It is a macrophotograph which shows the state which the gelatin gel which does not contain an insoluble residue melt | dissolves. A macroscopic photograph showing the state 2 hours after the dried gelatin gel that had been irradiated with ultraviolet rays at an integrated dose of 200 mJ / cm ² was rehydrated with PBS at 4 ° C and then transferred into an incubator at 37 ° C. is there. A macroscopic photograph showing the state 2 hours after the dried gelatin gel that had been irradiated with ultraviolet rays at an integrated dose of 400 mJ / cm ² was rehydrated with PBS at 4 ° C and then transferred into an incubator at 37 ° C. is there. A macroscopic photograph showing the state 2 hours after the dried gelatin gel that had been irradiated with ultraviolet rays at an integrated dose of 400 mJ / cm ² was rehydrated with PBS at 4 ° C and then transferred into an incubator at 37 ° C. is there. A macrophotograph showing the state 2 hours after the dried gelatin gel that had been irradiated with ultraviolet rays at a cumulative dose of 800 mJ / cm ² was rehydrated with PBS at 4 ° C and then transferred into an incubator at 37 ° C. is there. A macroscopic photograph showing the condition of the second day after the dried gelatin gel irradiated with ultraviolet rays at an accumulated dose of 800 mJ / cm ² is rehydrated with PBS at 4 ° C and then transferred to a 37 ° C incubator. is there. A macroscopic photograph showing the condition of the third day after the dried gelatin gel irradiated with ultraviolet rays at an accumulated dose of 800 mJ / cm ² is rehydrated with PBS at 4 ° C and then transferred to a 37 ° C incubator. is there. A macroscopic photograph showing the condition of the seventh day after the dried gelatin gel irradiated with ultraviolet rays at an accumulated dose of 800 mJ / cm ² is rehydrated with PBS at 4 ° C and then transferred to a 37 ° C incubator. is there. A macroscopic photograph showing the state 2 hours after the dried gelatin gel irradiated with ultraviolet rays at an integrated dose of 1,600 mJ / cm ² is rehydrated with PBS at 4 ° C and then transferred into a 37 ° C incubator. It is. Macroscopic photograph showing the condition of the seventh day after the dried gelatin gel irradiated with ultraviolet rays at an integrated dose of 1,600 mJ / cm ² is rehydrated with PBS at 4 ° C and then transferred into a 37 ° C incubator. It is. It is a macroscopic photograph showing a state in which a dried gelatin gel from which insoluble residues have been removed is irradiated with ultraviolet rays at an integrated dose of 800 mJ / cm ² and then rehydrated with PBS at 4 ° C. It is a macroscopic photograph showing a state where a dried gelatin gel from which insoluble residues have been removed is irradiated with ultraviolet rays at an integrated dose of 1,600 mJ / cm ² and then rehydrated with PBS at 4 ° C. Shows the state of the first day after the gelatin vitrigel produced through the process of irradiating with ultraviolet rays at an integrated dose of 800 mJ / cm ² (400 mJ / cm ² × 2 on one side) is placed in an incubator at 37 ° C. It is a macroscopic photograph. Shows the condition of the second day after the gelatin vitrigel prepared through the process of irradiating with ultraviolet rays at an integrated dose of 800 mJ / cm ² (400 mJ / cm ² × 2 on one side) is placed in an incubator at 37 ° C. It is a macroscopic photograph. Shows the state of the third day after the gelatin vitrigel prepared through the process of irradiating with ultraviolet rays at an integrated dose of 800 mJ / cm ² (400 mJ / cm ² × 2 on one side) is placed in an incubator at 37 ° C. It is a macroscopic photograph. 7th day after the gelatin vitrigel prepared through the process of irradiating ultraviolet rays with an integrated dose of 800 mJ / cm ² (400 mJ / cm ² × 2 on one side) is placed in a 37 ° C incubator. It is a macroscopic photograph. Shows the condition on the 7th day after gelatin vitrigel prepared through the process of irradiating ultraviolet rays at an integrated dose of 1,600 mJ / cm ² (800 mJ / cm ² × 2 on one side) in a 37 ° C incubator It is a macroscopic photograph. A macrophotograph showing a state in which gelatin vitrigel prepared through the process of irradiating with ultraviolet rays at an integrated dose of 1,600 mJ / cm ² (800 mJ / cm ² × 2 on one side) continued to be heated at 50 ° C for 24 hours. is there. A macroscopic photograph showing gelatin vitrigel heated for 3 minutes at 95 ° C after being irradiated with ultraviolet rays at an integrated dose of 1,600 mJ / cm ² (800 mJ / cm ² × 2 on one side) is there. Gelatin vitrigel prepared through the process of irradiating with ultraviolet rays at an integrated dose of 1,600 mJ / cm ² (800 mJ / cm ² × 2 on one side) was heated at 95 ° C for 3 minutes and then picked up with tweezers. It is a macroscopic photograph showing a state. Gelatin vitrigel prepared through the process of irradiating ultraviolet rays with an integrated dose of 1,600 mJ / cm ² (800 mJ / cm ² × 2 on one side) is cut to about 2.5 x 1.0 cm with a knife, and then edible oil at 180 ° C It is a macroscopic photograph showing a state of being fried and taken out for about 1 minute. Human dermis-derived fibroblasts are seeded on a tuna-derived gelatin vitrigel prepared through a process of irradiating with ultraviolet rays at an integrated dose of 1,600 mJ / cm ² (800 mJ / cm ² × 2 on one side) and cultured for 3 days. It is the microscope picture which observed the form of the cell which carried out the fluorescence dyeing | staining using calcein-AM and ethidium homodimer after the fluorescence microscope. Bar in the figure indicates 0.2 mm. Human dermis-derived fibroblasts are seeded on a tuna-derived gelatin vitrigel prepared through a process of irradiating with ultraviolet rays at an integrated dose of 1,600 mJ / cm ² (800 mJ / cm ² × 2 on one side) and cultured for 3 days. It is the microscope picture observed with the phase-contrast microscope after doing. Bar in the figure indicates 0.2 mm. It is a macroscopic photograph of a tuna-derived gelatin vitrigel film infiltrated with a rose extract solution for 5 minutes. It is a macroscopic photograph immediately after adsorbing the gelatin vitrigel membrane infiltrated with the rose extract solution on the human skin surface. 1 is a macroscopic photograph of a rose extract-containing gelatin vitrigel film in close contact with the human skin surface.

The method for producing gelatin vitrigel of the present invention is a method for producing gelatin vitrigel mainly composed of gelatin, and comprises the following steps:
(1) A step of producing a gelatin gel containing gelatin as a main component by cooling an aqueous gelatin solution containing insoluble collagen or collagen that can be refibrinated.
(2) A step of drying a gelatin gel to produce a vitrified gelatin gel dried body;
(3) a step of irradiating the dried gelatin gel with ultraviolet rays; and (4) a step of rehydrating the dried gelatin gel after the step (3) to prepare a gelatin vitrigel.

  Hereinafter, an embodiment of a method for producing a gelatin vitrigel of the present invention will be described along each of the above steps.

  In the step (1), a gelatin aqueous solution containing gelatin as a main component is prepared by cooling an aqueous gelatin solution containing insoluble collagen or collagen capable of refibrosis.

  The gelatin aqueous solution used in the step (1) can be insoluble collagen or refibrosis after the crude gelatin aqueous solution containing insoluble collagen as an insoluble residue derived from animal skin, or after dissolving purified gelatin in a solvent A mixed aqueous solution to which an aqueous solution containing collagen is added is preferable. Further, the mixed aqueous solution containing insoluble collagen or collagen capable of refibration is preferably a uniform aqueous solution to which a small amount of an unmodified acid-soluble collagen aqueous solution is added.

  As the gelatin, for example, a conventionally known one can be appropriately used depending on the application, and specifically, a gelatin derived from livestock products such as cattle, pigs and birds, and seafood such as seafood. It can be illustrated.

  In particular, in order to keep costs low, it is preferable to use a crude gelatin aqueous solution made from raw fish-derived hides. Here, the raw skin derived from fish includes fish skin and scales containing gelatin.

  When the raw skin derived from fish is used as a raw material, the type of fish is not particularly limited. Specifically, for example, fish such as tuna, tilapia, carp, Nile perch, salmon, scallop, tropical scallop, pollack, hake, mackerel can be used. From the viewpoint, tuna, tilapia and the like are particularly preferable. In particular, tuna is the most used fish species in Japan, and the amount used is 480,000 tons, including landing and import. However, only about 1% of the skin portion is effectively used except that about 50% is waste and the remaining about 50% is feed. Therefore, it is particularly preferable to use tuna-derived gelatin from the viewpoint of effective use of biomass.

  Examples of the insoluble residue contained in the crude extracted gelatin include insoluble collagen and hydroxyapatite. Here, in order to produce a gelatin vitrigel that does not dissolve even when heated, it is particularly necessary to use a gelatin aqueous solution containing a small amount of insoluble collagen and / or collagen that can be re-fibrinated. The gelatin content in the gelatin aqueous solution, and the amount of insoluble collagen and re-fibrible collagen can be appropriately adjusted in consideration of the thickness of the gelatin vitrigel to be produced.

  The ratio of the insoluble residue contained in the crude extracted gelatin varies depending on the raw material, the extraction method, and the like, but the ratio of the insoluble residue can be appropriately adjusted according to the use of the gelatin vitrigel. For example, when animal raw skin is shredded and then alkali-treated and gelatin is roughly extracted by heat treatment, and then large insolubles are removed by filtration, the insoluble residue is 0.1% to 20% of the volume of the gelatin aqueous solution. Can be about.

  Further, the ratio of insoluble collagen and re-fibrotic collagen to purified gelatin can be appropriately set according to the use of gelatin vitrigel. Specifically, the ratio of adding an unmodified 0.5 (w / v)% acid-soluble collagen aqueous solution (an aqueous solution containing collagen capable of refibrosis) to the purified gelatin aqueous solution is as follows: For example, a range of about 0.01 to 90%, preferably about 10 to 70% can be exemplified. When this value is converted as the content of insoluble collagen with respect to the purified gelatin aqueous solution, a range of about 0.005 to 0.45 (w / v)%, preferably about 0.05 to 0.35 (w / v) can be preferably exemplified. In other words, when the ratio of the aqueous solution containing collagen that can be refibrated to the purified gelatin aqueous solution or the content of insoluble collagen is within this range, the thermal stability of gelatin vitrigel is improved, so that medical materials, cosmetics, foods It can be used effectively industrially as a material.

  Solvents for dissolving purified gelatin powder and freeze-dried products include purified water, PBS (Phosphate Buffered Saline), physiological saline, HBSS (Hank's Balanced Salt Solution), basic culture solution, serum-free culture solution, or serum. Examples thereof include a culture broth. In particular, when gelatin is dissolved in a balanced salt solution such as PBS that retains physiological salt concentration and hydrogen ion concentration (pH), re-fibration of native collagen when mixed with an acid-soluble collagen aqueous solution. Can be effectively induced. On the other hand, when gelatin is dissolved in purified water or the like that is not a balanced salt solution, it is difficult to induce refibration of collagen. In this case, it is preferable to disperse insoluble collagen separately. In this way, a gelatin vitrigel that retains thermal stability can be produced even when purified gelatin is used.

  Then, a predetermined amount of a crude extracted gelatin aqueous solution containing insoluble collagen as an insoluble residue, or a gelatin aqueous solution containing a small amount of insoluble collagen or collagen that can be refibrinated is injected into a plastic culture dish, for example. The gelatin gel is produced by cooling to 20 ° C. or less for several hours to gelation. Generally, gelatin gelation is completed by cooling to 20 ° C. or more for several hours or more. However, at 4 ° C., gelatin gelation is completed by cooling for about 2 hours. The pH of the gelatin aqueous solution to be gelled is preferably about 6-8.

  Further, the gelatin concentration in the aqueous gelatin solution obtained by dissolving the crude extracted gelatin solution and the purified gelatin can be in the range of about 0.5 to 5.0%, preferably about 1.5 to 3.0%. If it is 0.5% or less, it is too dilute and gelation is weak, and if it is 5.0% or more, the concentration is too high to make uniforming difficult.

  In step (2), the gelatin gel is dried to produce a vitrified gelatin gel dried body.

  Various methods such as air drying, drying in an airtight container (circulating air in the container and always supplying dry air), and drying in an environment where silica gel is placed can be used to dry the gelatin gel. it can. For example, as an air-drying method, a method of drying for several hours to several days in an incubator kept sterile at 10 ° C. and 40% humidity, or a method of drying at room temperature of 20 ° C. or less overnight in an aseptic clean bench Etc. can be illustrated.

  By drying the gelatin gel, after the free water in the gelatin gel is removed, the bound water begins to be gradually removed from the gelatin and collagen molecules, whereby a dried gelatin gel can be prepared. As the period of the vitrification step is lengthened, a gelatin vitrigel having excellent transparency and strength when rehydrated can be obtained.

  In the step (3), the dried gelatin gel is irradiated with ultraviolet rays.

  By irradiating the dried gelatin gel with ultraviolet rays, the thermal stability of the gelatin vitrigel can be improved. This is thought to be due to the introduction of cross-links to the dried gelatin gel by ultraviolet irradiation.

The irradiation amount of ultraviolet rays to the dried gelatin gel can be appropriately set according to the size and thickness of the dried gelatin gel, for example, 400 mJ / cm ^{2 to} 2000 mJ / cm ² in terms of the integrated irradiation amount, Preferably, it can be 800 mJ / cm ² or more.

Moreover, it is preferable to irradiate the gelatin gel dried product with ultraviolet rays on both surfaces (front and back surfaces) of the gelatin gel dried product. In this case, it is preferable that the amount of ultraviolet light irradiated on both sides of the dried gelatin gel is 800 mJ / cm ² or more for each side of the dried gelatin gel. Thereby, the thermal stability of gelatin vitrigel can be improved.

  As will be described later, the dried gelatin gel can be used effectively industrially because a gelatin vitrigel having thermal stability can be produced by rehydration when necessary.

  In step (4), the dried gelatin gel obtained in step (3) is rehydrated to produce gelatin vitrigel.

  A gelatin vitrigel can be prepared by rehydrating a dried gelatin gel with purified water, PBS, culture solution or the like.

  The liquid to be rehydrated may contain various components according to the use of gelatin vitrigel.

  Specifically, when using gelatin vitrigel as a medical material, for example, various drugs including antibiotics, cell growth factor, differentiation inducing factor, cell adhesion factor, antibody, enzyme, cytokine, hormone, A lectin or a physiologically active substance such as fibronectin, vitronectin, entactin, or osteopoietin may be contained as an extracellular matrix component that does not gel.

  Since gelatin vitrigel containing a physiologically active substance can supply factors necessary for cell adhesion, proliferation, differentiation and the like from the gelatin vitrigel side, a better culture environment can be realized. Moreover, it is very useful for conducting a test for examining the effect of the contained physiologically active substance on cells. In addition, gelatin vitrigel containing a physiologically active substance can function as a drug delivery system by being implanted into the body.

  Furthermore, gelatin vitrigel can permeate a physiologically active substance having a large molecular weight, thereby allowing mutual interaction between each cell seeded on the front and back surfaces of this gelatin vitrigel via the physiologically active substance. Can greatly contribute to testing and research of action.

  Gelatin vitrigel can also be used as a cell culture carrier for culturing cells. Examples of cells to be cultured include primary cultured cells, established cells, fertilized eggs, and cells obtained by introducing a foreign gene into these cells. Furthermore, these cells may be undifferentiated stem cells, cells in the process of differentiation, terminally differentiated cells, and dedifferentiated cells. Moreover, as means for initiating the culture of these cells, cell suspension, shredded tissue pieces, fertilized eggs, or seeding of three-dimensionally reconstructed gel cultures or multicellular aggregates (spheroids) can be mentioned. . That is, adherent cells that can be cultured by existing methods can be cultured on gelatin vitrigel.

  Furthermore, for example, one or more types of cells can be cultured on both sides of the above-mentioned cells as well as one side of gelatin vitrigel. In double-sided culture, it is possible to culture different types of cells on each side. In particular, by culturing epithelial cells on one side of the gelatin vitrigel and mesenchymal cells on the other side, a percutaneous absorption model or intestinal absorption model with epithelial-mesenchymal interaction, etc. By culturing vascular endothelial cells on one surface and cancer cells on the other surface, cell function assays such as an angiogenesis model and a cancer invasion model are made possible.

  The gelatin vitrigel of the present invention is excellent in thermal stability and, for example, does not dissolve even at a living body temperature (approximately 37 ° C.), and thus can be a medical material that can be effectively used particularly in the field of regenerative medicine.

  In addition, when gelatin vitrigel is used as a cosmetic product, for example, it can contain any component normally used in cosmetics. Such optional ingredients include, for example, macadamia nut oil, avocado oil, corn oil, olive oil, rapeseed oil, sesame oil, castor oil, safflower oil, cottonseed oil, jojoba oil, palm oil, palm oil, mint oil, liquid lanolin, Oils such as hardened coconut oil, hardened oil, mole, hardened castor oil, beeswax, candelilla wax, carnauba wax, ibotarou, lanolin, reduced lanolin, hard lanolin, jojoba wax, waxes, liquid paraffin, squalane, pristane, ozokerite, paraffin, Hydrocarbons such as ceresin, petrolatum, microcrystalline wax, higher fatty acids such as oleic acid, isostearic acid, lauric acid, myristic acid, palmitic acid, stearic acid, behenic acid, undecylenic acid, cetyl alcohol, stearyl alcohol, Higher alcohols such as sostearyl alcohol, behenyl alcohol, octyldodecanol, myristyl alcohol, cetostearyl alcohol, cetyl isooctanoate, isopropyl myristate, hexyldecyl isostearate, diisopropyl adipate, di-2-ethylhexyl sebacate, cetyl lactate, Synthetic ester oils such as diisostearyl malate, ethylene glycol di-2-ethylhexanoate, neopentyl glycol dicaprate, glycerin di-2-heptylundecanoate, pentane erythritol tetra-2-ethylhexanoate, dimethylpoly Chain polysiloxane such as siloxane, methylphenyl polysiloxane, diphenyl polysiloxane, octamethylcyclotetrasiloxane, decamethylcyclopentadi Oils such as silicone oils such as cyclopolysiloxanes such as xane and dodecamethylcyclohexanesiloxane, amino-modified polysiloxanes, polyether-modified polysiloxanes, alkyl-modified polysiloxanes and fluorine-modified polysiloxanes, fatty acid soaps (laurin) Sodium sulfate, sodium palmitate, etc.), anionic surfactants such as potassium lauryl sulfate, alkylsulfuric triethanolamine ether, cationic surfactants such as stearyltrimethylammonium chloride, benzalkonium chloride, laurylamine oxide, imidazoline series Amphoteric surfactants (such as 2-cocoyl-2-imidazolinium hydroxide-1-carboxyethyloxy disodium salt), betaine surfactants (alkylbetaines, amidebetas) In, sulfobetaine, etc.), amphoteric surfactants such as acylmethyltaurine, polyethylene glycol, glycerin, 1,3-butylene glycol, erythritol, sorbitol, xylitol, maltitol, propylene glycol, dipropylene glycol, diglycerin, isoprene Moisturizing of polyhydric alcohols such as glycol, 1,2-pentanediol, 2,4-hexylene glycol, 1,2-hexanediol, 1,2-octanediol, sodium pyrrolidonecarboxylate, lactic acid, sodium lactate, etc. Classification, guar gum, quince seed, carrageenan, galactan, gum arabic, pectin, mannan, starch, xanthan gum, curdlan, methylcellulose, hydroxyethylcellulose, carboxymethylcellulose, Cyl hydroxypropyl cellulose, chondroitin sulfate, dermatan sulfate, glycogen, heparan sulfate, hyaluronic acid, sodium hyaluronate, tragacanth gum, keratan sulfate, chondroitin, mucoitin sulfate, hydroxyethyl guar gum, carboxymethyl guar gum, dextran, kerato sulfate, locust bean gum, Thickeners such as succinoglucan, caronic acid, chitin, chitosan, carboxymethylchitin, polyvinyl alcohol, polyvinylpyrrolidone, carboxyvinyl polymer, sodium polyacrylate, polyethylene glycol, bentonite, paraaminobenzoic acid UV absorber, anthranilic acid UV absorber, salicylic acid UV absorber, cinnamic acid UV absorber, benzophenone UV absorber, sugar UV Absorbers, ultraviolet absorbers such as 2- (2′-hydroxy-5′-t-octylphenyl) benzotriazole, 4-methoxy-4′-t-butyldibenzoylmethane, lower alcohols such as ethanol and isopropanol , Vitamin A or its derivatives, Vitamin B6 hydrochloride, Vitamin B6 tripalmitate, Vitamin B6 dioctanoate, Vitamin B2 or its derivatives, Vitamin B such as Vitamin B12, Vitamin B15 or its derivatives, Vitamin E such as Vitamin E acetate Vitamins such as vitamin D, vitamin H, pantothenic acid, panthetin, pyrroloquinoline quinone and the like can be preferably exemplified.

  The gelatin vitrigel of the present invention is excellent in thermal stability even at a living body temperature (approximately 37 ° C.), for example, and can be prepared so as not to dissolve at all or can be prepared so as to dissolve gradually. It can be effectively used not only for medical materials but also for cosmetics such as cosmetics and fragrances.

  Furthermore, when gelatin vitrigel is used as a food material, for example, sugar and sugar alcohols, meat and fish extracts, proteins, peptides, amino acids, dietary fiber, vitamins, starches, dextrin, fats and oils, Components such as alcohols, salts, seasonings, spices, preservatives, sweeteners, colorants, quality improvers, and fragrances can be included.

  The gelatin vitrigel of the present invention is excellent in thermal stability and, for example, can be kept in a solid form without being dissolved by boiling water at about 100 ° C. or oil at 180 ° C. or higher. Therefore, the gelatin vitrigel of the present invention can be a completely new food material that can maintain the texture and taste even when cooked.

  Furthermore, as a step (5), the gelatin vitrigel produced by the method including the above steps (1) to (4) is re-dried and vitrified, thereby producing a dried gelatin vitrigel. be able to. This dried gelatin vitrigel can be converted to gelatin vitrigel again by rehydration when necessary, so that it can be effectively used industrially.

Furthermore, in the production of the dried gelatin vitrigel, the step (6) may include a step of irradiating the dried gelatin vitrigel produced in the step (5) with ultraviolet rays. For example, in step (3), it is difficult to irradiate ultraviolet rays on both sides of the dried gelatin gel when it is difficult to reverse the dried gelatin gel due to adhesion of the dried gelatin gel to a culture dish or the like. There can be. In such a case, for example, in step (3), one side (one side) of the dried gelatin gel is irradiated with a desired amount of ultraviolet light, and in step (4), the dried gelatin gel is re-watered. A gelatin vitrigel is prepared by adding the gelatin vitrigel from the culture petri dish and the like, and after reversing the front and back, drying is again performed in step (5) to obtain a dried vitrified gelatin vitrigel. In the step (6), the other surface of the dried gelatin gel (the surface not irradiated with the ultraviolet ray in the step (3)) can be irradiated with ultraviolet rays. This gelatin vitrigel dried product can be rehydrated again to produce a gelatin vitrigel. As described above, the gelatin vitrigel that has undergone such a process is irradiated with ultraviolet rays from both sides (front and back surfaces) during the production process, and thus has improved thermal stability. Also in this case, it is preferable that the irradiation amount of the ultraviolet rays is 800 mJ / cm ² or more for each side of the dried gelatin gel and the dried gelatin vitrigel.

  The gelatin vitrigel of the present invention and the method for producing the same are not limited to the above forms. For example, the forms proposed by the present inventors so far (Patent Documents 1, 2, PCT / JP2011 / 069191) Can be taken into account.

  Specifically, for example, a support made of a nylon membrane or the like can be introduced into the gelatin aqueous solution described above. In addition, gelation, vitrification, and rehydration can be performed by injecting gelatin dissolved (solated) into the inside using a wall surface mold placed on a substrate (PCT / JP2011 / 069191 ). Furthermore, the gelatin aqueous solution described above is layered on the dried gelatin gel to be gelled, vitrified and rehydrated (PCT / JP2011 / 069191), gelatin vitrigel already prepared in the gelatin aqueous solution described above or its dried product Gelatin vitrigels of any thickness can be made by embedding the body and gelling, vitrifying and rehydrating. Moreover, it can also vitrify on the film which a gelatin gel dried body or a gelatin vitrigel dried body can peel (PCT / JP2011 / 069191). In this case, the handleability of the dried gelatin gel is improved, and the dried gelatin gel can be easily irradiated with ultraviolet rays.

  EXAMPLES Hereinafter, although an Example demonstrates this invention further in detail, this invention is not limited to these Examples at all.

[Example 1: Gelatin vitrigel prepared from sashimi collagen slice (Eco & Health Lab product number 80924)]
Sashimi Collagen Slice (Eco & Health Labs product number 80924), which is commercially available as a frozen product, cuts the tuna skin into small pieces, then roughly extracts gelatin by alkali treatment and heat treatment, and then removes large insoluble matter by filtration. It is a gelatin gel mainly composed of tuna gelatin prepared by removing. Since this gelatin gel is prepared by the process described above, it can be easily converted into a solution state by heating to about 40 ° C., but this gelatin solution contains small insoluble residues (FIG. 1).

  2 ml of the gelatin solution solubilized at 40 ° C. was poured into a petri dish for cell culture (FALCON Cat. No353001) having a diameter of 35 mm and allowed to stand at 4 ° C. for 2 hours for gelation. By leaving this gelatin gel in an incubator at a temperature of 10 ° C and a humidity of 40% for 2 days with the lid removed, the free water in the gelatin gel is completely removed by the 2nd day, resulting in a dry state. I found out that Here, vitrification has progressed from the point where free water has been completely removed. Therefore, according to the conventional method for producing vitrigel, the dried body (gelatin gel dried body) in a dry state can be rehydrated. Gelatin vitrigel can be prepared. Therefore, the free water was completely removed, and 2 ml of PBS was added to the culture petri dish where vitrification had begun at 4 ° C for rehydration. This gelatin vitrigel is adsorbed on the bottom surface of the culture dish, and can be peeled off by tweezers after being scratched on the bottom side with the tip of the tweezers along the wall of the culture dish (FIG. 2). The exfoliated gelatin vitrigel was brittle and difficult to handle (FIG. 3). Also, from the viewpoint of applying this gelatin vitrigel as a regenerative medicine material, it was transferred into an incubator at 37 ° C, and as a result of observation over time, almost everything was dissolved in PBS within 2 hours. It was found that it could not be maintained (FIG. 4).

[Comparative Example 1: Examination of preparation of vitrigel from gelatin of Example 1 with insoluble residue removed]
The insoluble small residue present in the gelatin solution of Example 1 is obtained by treating the gelatin solution with a centrifuge [3000 rpm (= 1400 g) × 30 minutes]] and filtering the supernatant with a 0.45 μm filter. It was able to be removed (FIG. 5). Using the gelatin solution from which this residue was removed, an attempt was made to produce gelatin vitrigel in the same manner as in Example 1. As a result, when 2 ml of PBS was poured into the culture dish at 4 ° C, almost all of the dried gelatin gel without residue, which was in a dry state, was dissolved in PBS and recovered as gelatin vitrigel. It was not possible (FIG. 6). This suggested that preparation of gelatin vitrigel requires a small insoluble residue in the gelatin solution. Next, since this insoluble residue can be recovered as a precipitate after the above-mentioned centrifugation, the characteristics thereof were analyzed. As a result, it was found that this precipitate was easily dissolved in dilute hydrochloric acid. Considering the production process of the sashimi collagen slice described in Example 1, this precipitate (insoluble residue in gelatin solution) was dissolved in dilute hydrochloric acid, so that it was presumed to be insoluble collagen.

[Example 2: Gelatin vitrigel excellent in thermal stability and handleability]
In the same manner as in Example 1, 2 ml of gelatin solution solubilized at 40 ° C was poured into a petri dish for cell culture having a diameter of 35 mm, covered and allowed to stand at 4 ° C for 2 hours to gel. The gelatin gel was left in an incubator at a temperature of 10 ° C and a humidity of 40% for 2 days with the lid removed to completely remove the free water in the gelatin gel and proceed to vitrification, thereby bringing it into a dry state ( Gelatin gel dried body). Next, after removing the lid of the cell culture petri dish, the dried gelatin gel in a dry state is FUNA-UV LINKER (product code FS-1500, manufactured by Funakoshi Co., Ltd.). Ultraviolet rays were irradiated so as to be 800 and 1,600 mJ / cm ² . After irradiating with ultraviolet rays at each accumulated dose, it was peeled off by adding 2 ml of PBS at 4 ° C. in a petri dish for cell culture in the same manner as in Example 1 to rehydrate. In this peeling process, the handling property of gelatin vitrigel irradiated with ultraviolet rays at an integrated dose of 200 mJ / cm ² was almost the same as the handling property of gelatin vitrigel of Example 1 not irradiated with ultraviolet rays, but it was brittle. The handleability of gelatin vitrigel irradiated with ultraviolet rays at an integrated dose of 400 mJ / cm ² or more improved with an increase in the integrated dose. The gelatin vitrigel irradiated with ultraviolet rays was transferred into an incubator at 37 ° C. and observed over time. As a result, it was found that almost all gelatin vitrigel irradiated with ultraviolet rays at an integrated dose of 200 mJ / cm ² was dissolved in PBS within 2 hours and could not be maintained as gelatin vitrigel (FIG. 7). . Gelatin vitrigel irradiated with ultraviolet rays at an integrated dose of 400 mJ / cm ² was confirmed as a film-like form at 2 hours (Fig. 8), but almost all was in PBS at 24 hours. (FIG. 9). Gelatin vitrigel irradiated with ultraviolet rays at an accumulated dose of 800 mJ / cm ² was confirmed as a film-like form on the 1st, 2nd and 3rd days (FIGS. 10, 11, and 12). ) On the seventh day, although a film-like form was confirmed, the thickness was considerably thin (FIG. 13). On the other hand, gelatin vitrigel irradiated with ultraviolet rays at an integrated dose of 1,600 mJ / cm ² was confirmed to change into a film-like form with its surroundings turned up at the second hour (Fig. 14). It was confirmed that the same form was maintained at the eye point (FIG. 15).

[Comparative Example 2: Examination of preparation of vitrigel from gelatin of Example 2 from which insoluble residue was removed]
Comparative Example 1 and using a gelatin solution that remove small insoluble residue present in the gelatin solution of Example 1 in the same manner, in the same manner as in Example 2 800 mJ / cm ² and 1,600mJ / cm ² Preparation of gelatin vitrigel was attempted after irradiating ultraviolet rays at an integrated dose.

  As a result, as in Comparative Example 1, when 2 ml of PBS was poured into the culture dish at 4 ° C., almost all the dried gelatin gel without residue that had been in a solidified state was dissolved in PBS. However, it could not be recovered as gelatin vitrigel (FIGS. 16 and 17).

[Example 3: Gelatin vitrigel excellent in shape stability, thermal stability and handleability]
In the same manner as in Example 2, dried gelatin gels derived from 2 ml of gelatin were prepared in a cell culture dish having a diameter of 35 mm, and the cumulative irradiation doses were 400 mJ / cm ² and 800 mJ / cm ^2. As shown in FIG. In this petri dish for cell culture, add 2 ml of PBS at 4 ° C and rehydrate to peel off the gelatinous vitrigel in the form of a film, and reverse it to the bottom of the newly prepared cell culture dish. did. With the lid of the cell culture petri dish removed, leave it in an incubator with a temperature of 10 ° C and a humidity of 40% for 2 days to completely remove the free water in the inverted gelatin vitrigel and vitrify it again. To advance to dryness (dried gelatin vitrigel). Next, after removing the lid of the petri dish for cell culture, the dried gelatin vitrigel in the dried state was irradiated with an accumulated dose of 400 mJ / cm ² last time. cm ² become as, also such that the last similar 800 mJ / cm ² in gelatin Vito Rigel dried body was irradiated in the previous 800 mJ / cm ^2, the total irradiation amount of each surface by irradiation of ultraviolet light is 800 mJ / Ultraviolet rays were irradiated so as to be cm ² (400 mJ / cm ² × 2 on one side) and 1,600 mJ / cm ² (800 mJ / cm ² × 2 on one side). Thereafter, as in Example 2, 2 ml of PBS was added to the cell culture petri dish at 4 ° C. and rehydrated. As a result, the gelatinous vitrigel in the form of a film could be peeled from any cell culture petri dish. . After irradiating these both surfaces with ultraviolet rays, gelatin vitrigel recovered as a film-like form was transferred into a 37 ° C. incubator and observed over time. As a result, gelatin vitrigel irradiated with ultraviolet rays at an integrated dose of 800 mJ / cm ² (400 mJ / cm ² × 2 on one side) is in the form of a film on the first, second and third days. Although it was confirmed (FIGS. 18, 19, and 20), the thickness gradually decreased and dissolved in PBS until the film-like morphology could not be confirmed at the 7th day (FIG. 21). On the other hand, gelatin vitrigel irradiated with ultraviolet rays at an integrated dose of 1,600 mJ / cm ² (800 mJ / cm ² × 2 on one side) does not change its shape at 37 ° C. It was confirmed that the film-like form immediately after peeling was maintained at the time (FIG. 22).

In addition, the gelatin vitrigel irradiated with ultraviolet rays with the integrated irradiation dose of 1,600 mJ / cm ² (800 mJ / cm ² × 2 on one side) described above continued to warm for 24 hours by raising the temperature to 50 ° C. After that, it was confirmed that the film-like shape immediately after peeling was maintained with almost no change in shape (FIG. 23). Furthermore, when the temperature was raised to 95 ° C. and kept warming for 3 minutes, the peripheral portion changed into a slightly shrunken shape (FIG. 24), but this gelatin vitrigel film was picked with tweezers without melting or breaking. It was confirmed that it was raised (FIG. 25). Therefore, wipe off excess water around the gelatin vitrigel membrane rehydrated at room temperature, cut the gelatin vitrigel membrane to about 2.5 x 1.0 cm with a knife, and then float it in cooking oil prepared at 180 ° C. I fried about 1 minute until the sound calmed down. As a result, it was confirmed that the gelatin vitrigel film changed to a slightly atrophied form without dissolving (FIG. 26).

  The gelatin vitrigel of the present invention has been confirmed to be effective as, for example, a new food material because it has excellent thermal stability and can maintain its shape without being dissolved even at high temperatures.

[Example 4: Culture of human dermis-derived fibroblasts on gelatin vitrigel excellent in shape stability, thermal stability and handling]
On the tuna-derived gelatin vitrigel membrane (processed from sashimi collagen slice) prepared through the step of irradiating ultraviolet rays with an integrated dose of 1,600 mJ / cm ² (800 mJ / cm ² × 2 on one side) in Example 3 above Then, human dermis-derived fibroblasts were seeded and cultured for 3 days, and the morphology of the cells was observed by phase contrast microscopy. Thereafter, cell viability was determined by fluorescent staining using calcein-AM and ethidium homodimer. As a result, it was found that most of the human dermis-derived fibroblasts survived on the tuna-derived gelatin vitrigel membrane (FIG. 27) and exhibited a good spreading form (FIG. 28). In other words, it was suggested that the tuna-derived gelatin vitrigel film is safe without cytotoxicity.

[Example 5: Characteristics of vitrigel prepared from gelatin (RM-100 manufactured by Zerais Co., Ltd.) and native collagen (I-AC manufactured by Koken Co., Ltd.)]
In Example 2 described above, when a gelatin solution containing tuna gelatin as a main component was used, gelatin vitrigel prepared through the step of irradiating with ultraviolet rays at an accumulated dose of 800 mJ / cm ² was obtained on the first and second days. It was confirmed that it was maintained as a membranous form at the time of eye and day 3. However, in Comparative Example 2, when the gelatin solution from which the insoluble small residue present in the gelatin solution is removed is used, it can be produced through a process of irradiating with ultraviolet rays at an integrated dose of 800 mJ / cm ² or more. It was confirmed that gelatin vitrigel could not be prepared. In Comparative Example 1, it was assumed that the insoluble small residue present in the gelatin solution was insoluble collagen because it was dissolved in dilute hydrochloric acid.

Based on the above background, in this example, after gelatinization was evaluated by allowing a mixed solution obtained by mixing a purified gelatin solution to a purified gelatin solution to stand at 4 ° C. for 2 hours, a temperature of 10 ° C. and a humidity of 40 The purpose of this study is to investigate whether vitrigel stable at 37 ° C can be prepared by irradiating with UV at an integrated dose of 800 mJ / cm ² and rehydrating with PBS after vitrification in an incubator It was. Specifically, purified porcine gelatin (RM-100 manufactured by Zerais) was dissolved in purified water or PBS heated to 40 ° C. so that the final concentration was 2.5 (w / v)%, and then 2.5% 0.5% bovine-derived native collagen solution (I-AC manufactured by Koken Co., Ltd.) is added to the gelatin solution at volume ratios of 10: 0, 9: 1, 8: 2, 7: 3, 5: 5, 3: 7, and 1: When the above-mentioned gelatin gelation and vitrigel preparation were studied using the mixed solution mixed so as to be 9, the results shown in Table 1 were obtained.

  Specifically, in the case of a gelatin solution containing no collagen solution, the gelatin gel could be prepared by cooling at 4 ° C. regardless of the solvent, but when the gelatinized dried gelatin gel was further rehydrated with PBS. In this case, it was dissolved in PBS and vitrigel could not be prepared.

  When purified water is used as a solvent for dissolving gelatin, <1> Gelatin gelation is not induced even when cooled to 4 ° C in a mixed solution containing 70% or more collagen solution. When rehydrated, a stable vitrigel can be prepared at 4 ° C or 37 ° C. <2> Gelatin gel can be prepared by cooling at 4 ° C in a mixed solution containing 10% to 50% collagen solution. Vitrigel can be prepared at 4 ° C even when rehydrated with PBS, but when heated to 37 ° C, it can dissolve in PBS within 1 hour except for vitrigel derived from a mixture containing 50% collagen solution. I understood. Here, the reason why a vitrigel stable at 37 ° C. was prepared from an admixture containing 50% or more of a collagen solution is that a gel that has strength when vitrified collagen molecules are rehydrated in PBS. It was thought to be caused by fibrosis in the state.

  Furthermore, when the solvent that dissolves gelatin is PBS, gelation of gelatin is not induced even when cooled to 4 ° C in a mixture containing <1> collagen solution at 90% or more. When hydrated, a stable vitrigel can be produced at 4 ° C or 37 ° C. <2> Gelatin gel can be prepared by cooling at 4 ° C in a mixed solution containing 10% to 70% collagen solution. It was found that when rehydrated with PBS, a stable vitrigel was prepared at 4 ° C or 37 ° C. Here, the reason why a vitrigel stable at 37 ° C. could be prepared by vitrification and rehydration after gelatin gel was prepared from a mixture containing 10% to 70% collagen solution was as follows: This was thought to be due to the gradual fibrosis of the collagen molecules in PBS, a solvent in which gelatin was dissolved.

  From the above results, it is possible to prepare a gel mainly composed of gelatin from a mixture containing 10% collagen solution using PBS instead of purified water as a solvent for dissolving gelatin, and then vitrify and rehydrate. It was found that vitrigel composed mainly of gelatin stable at 37 ° C. can be produced.

  The gelatin vitrigel of the present invention can be made from a mixed aqueous solution obtained by adding an insoluble collagen or an aqueous solution containing collagen that can be refibrinated to purified purified gelatin, and has excellent thermal stability, even at about 37 ° C. Since it does not dissolve, for example, it was confirmed that it can be effectively used as a medical material by appropriately adding various physiologically active substances.

[Example 6: Usefulness of gelatin vitrigel as a cosmetic base material]
A tuna-derived gelatin vitrigel film (processed from a sashimi collagen slice) produced through the process of irradiating ultraviolet rays at an integrated dose of 1,600 mJ / cm ² (800 mJ / cm ² × 2 on one side) in Example 3 above. Then, it was re-glassed on a vinyl sheet to prepare a dried gelatinous vitrigel body. This dried gelatin vitrigel was infiltrated with mint water or rose extract solution (Nichirei Rose Extract Benikahime, Nichirei Bioscience Co., Ltd.) for 5 minutes (FIG. 29), and the adsorptivity to human skin was evaluated. The mint water was prepared by injecting 5 mL of WATER (SIGMA, W3500) into a 50 mL conical tube, adding 40 μl of mint oil (Kenei Pharmaceutical Co., Ltd.) and mixing well.

  As a result, the membranous gelatin vitrigel rehydrated by infiltration into mint water and rose extract solution could be easily adsorbed to human skin (FIG. 30). Further, it was found that the adsorbed gelatin vitrigel was gradually dried on the skin surface, but kept the fragrance for several hours or more (FIG. 31). Further, the dried gelatin vitrigel dried and adhered to the skin surface could be easily peeled off without feeling uncomfortable such as pain.

  It has been confirmed that the gelatin vitrigel of the present invention is excellent in thermal stability and can be effectively used as a cosmetic product by containing various cosmetic ingredients.

Claims (27)

A method for producing gelatin vitrigel comprising gelatin as a main component, the following steps:
(1) A step of producing a gelatin gel containing gelatin as a main component by cooling an aqueous gelatin solution containing insoluble collagen or collagen that can be refibrinated.
(2) A step of drying a gelatin gel to produce a vitrified gelatin gel dried body;
(3) a step of irradiating the dried gelatin gel with ultraviolet rays; and (4) a step of rehydrating the dried gelatin gel after the step (3) to produce a gelatin vitrigel. Vitrigel production method.   The method for producing a gelatin vitrigel according to claim 1, wherein in the step (3), both sides of the dried gelatin gel are irradiated with ultraviolet rays. 3. The method for producing a gelatin vitrigel according to claim 2, wherein the amount of ultraviolet rays applied to both sides of the dried gelatin gel is 800 mJ / cm ² or more for each side of the dried gelatin gel.   The method for producing a gelatin vitrigel according to any one of claims 1 to 3, wherein the gelatin aqueous solution in the step (1) is a crudely extracted gelatin aqueous solution containing insoluble collagen as an insoluble residue derived from an animal hide. .   The aqueous gelatin solution in the step (1) is a mixed aqueous solution to which an aqueous solution containing insoluble collagen or collagen capable of refibrosis is added after dissolving purified gelatin in a solvent. A method for producing any gelatin vitrigel.   6. The method for producing gelatin vitrigel according to claim 5, wherein the solvent for dissolving the purified gelatin is an equilibrium salt solution that maintains a physiological salt concentration and a hydrogen ion concentration (pH).   A gelatin vitrigel produced by the method according to claim 1.   A gelatin vitrigel produced by the method of claim 4, wherein the raw skin of the animal used as the raw material of the crude extracted gelatin aqueous solution is derived from fish.   The gelatin vitrigel according to claim 8, wherein the fish is a tuna.   A medical material comprising gelatin vitrigel produced by the method of claim 5.   A cosmetic comprising the gelatin vitrigel according to claim 7 and a cosmetic ingredient.   A food material comprising the gelatin vitrigel according to any one of claims 7 to 9 as a raw material. A method for producing a dried gelatin gel comprising gelatin as a main component and capable of producing a gelatin vitrigel, comprising the following steps:
(1) A step of producing a gelatin gel containing gelatin as a main component by cooling an aqueous gelatin solution containing insoluble collagen or collagen that can be refibrinated.
(2) A process for producing a dried gelatin gel by drying the gelatin gel; and (3) a method for producing a dried gelatin gel, comprising the step of irradiating the dried gelatin gel with ultraviolet rays.   14. The method for producing a dried gelatin gel product according to claim 13, wherein the gelatin aqueous solution in the step (1) is a crude extracted gelatin aqueous solution containing insoluble collagen as an insoluble residue derived from an animal hide.   14. The gelatin gel according to claim 13, wherein the gelatin aqueous solution in the step (1) is a mixed aqueous solution to which an aqueous solution containing insoluble collagen or collagen capable of refibrosis is added after dissolving purified gelatin in a solvent. A method for producing a dried body.   A dried gelatin gel produced by the method according to claim 13.   15. A dried gelatin gel produced by the method according to claim 14, wherein the raw hide of the animal used as the raw material for the crude extracted gelatin aqueous solution is derived from fish.   The dried gelatin gel according to claim 17, wherein the fish is tuna. A method for producing a dried gelatin vitrigel comprising gelatin as a main component, the following steps:
(1) A step of producing a gelatin gel containing gelatin as a main component by cooling an aqueous gelatin solution containing insoluble collagen or collagen that can be refibrinated.
(2) A step of drying a gelatin gel to produce a vitrified gelatin gel dried body;
(3) A step of irradiating the dried gelatin gel with ultraviolet rays;
(4) A step of rehydrating the gelatin gel dried material obtained through the step (3) to prepare a gelatin vitrigel; and (5) Redrying the gelatin vitrigel to produce a vitrified gelatin vitrigel dried material. The manufacturing method of the gelatin vitrigel dry body characterized by including this process.   The method for producing a dried gelatin vitrigel according to claim 19, further comprising the step of (6) irradiating the dried gelatin vitrigel with ultraviolet rays.   21. The ultraviolet ray is irradiated on one surface of the dried gelatin gel in the step (3), and the ultraviolet ray is irradiated on the other surface of the dried gelatin vitrigel in the step (6). Of producing a dried gelatin vitrigel product.   22. The dried gelatin vitrigel according to any one of claims 19 to 21, wherein the gelatin aqueous solution in the step (1) is a crude extracted gelatin aqueous solution containing insoluble collagen as an insoluble residue derived from an animal hide. Production method.   The aqueous gelatin solution in the step (1) is an aqueous solution to which an aqueous solution containing insoluble collagen or collagen capable of refibrosis is added after dissolving purified gelatin in a solvent. A method for producing a dried gelatin vitrigel.   24. A dried gelatin vitrigel produced by the method according to claim 19.   23. A dried gelatin vitrigel produced by the method according to claim 22, wherein the raw hide of the animal used as the raw material of the crude extracted gelatin aqueous solution is derived from fish.   The dried gelatin vitrigel according to claim 25, wherein the fish is tuna.   27. A gelatin vitrigel obtained by rehydrating the dried gelatin vitrigel according to any one of claims 24 to 26.