JP2008110207A - Infusion material to living body and cosmetic/medical bulk material - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an infusion material to a living body which is safe to infuse in a living body, is hardly absorbed in the living body and remains in an infused region for a long period of time, and also to provide a cosmetic/medical bulk material. <P>SOLUTION: The infusion material to a living body comprises a complex gel containing chitosans and collagen. The cosmetic/medical bulk material comprises a porous body containing the chitosans and collagen. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

  The present invention relates to a biomaterial, and more particularly, to a bioinjection material to be injected into a living body, and a cosmetic / medical bulk material for transplantation into the living body.

  In recent years, there is a great demand for biomaterials. For example, biomaterials to be injected into the subcutaneous tissue for the purpose of wrinkle stretching, congenital abnormalities, acquired abnormalities, compensation of body defects due to diseases, etc., cosmetic purposes such as enlargement of the chest and buttocks There is a need for biomaterials to do this. However, there are very few biomaterials that can be safely injected or inserted into the living body.

  Currently, collagen and hyaluronic acid are generally used as materials to be injected into a living body.

  Collagen has good biocompatibility and is often used as a cell scaffolding material. However, animal-derived collagen is derived from cattle and pigs and has unique properties depending on the type of animal, so it has rejection and allergic reactions. Problems that are likely to occur and infectious diseases common to humans and animals are problems. There is also a problem that it is absorbed very quickly in the living body.

  On the other hand, although hyaluronic acid is a synthetic product and high in safety, there is a problem that it is absorbed several months to half a year after injection. Therefore, in order to slow down the decomposition rate, “Super Hyaluronic Acid”, “Derma Live”, “Aquamid”, etc., in which a non-absorbable hydrogel is mixed have been developed. However, the particulate hydrogel cannot be repaired when it penetrates into the skin or dermis and causes tissue deformation in the body, and safety is a problem.

  Silicone is generally used as a material to be inserted into a living body. However, when silicone leaks locally, if the silicone particles penetrate into the skin and dermis and cause tissue deformation in the body, it cannot be repaired and safety is a problem.

  Therefore, as biomaterials, for example, implants for intradermal injection using biologically resorbable polymer microspheres or microparticles suspended in a gel (see Patent Document 1) and collagen derived from aquatic animals. A carrier and a biomaterial obtained by using (see Patent Document 2) have been proposed. However, these also have a problem of being fast absorbed by the living body.

JP 2000-516839 A (Claims etc.) Japanese translation of PCT publication No. 2003-534858 (Claims etc.)

  In view of such circumstances, the present invention is safe even when injected into a living body, is difficult to be absorbed by the living body, and can maintain the form at the injection site or transplant site over a long period of time, and The objective is to provide bulk materials for beauty and medical treatment.

  A first aspect of the present invention that solves the above problems is a living body injection material comprising a composite gel containing chitosans and collagen.

  In the first aspect, the combination of collagen having excellent biocompatibility and chitosans which are relatively hardly absorbed in the living body and have the property of easily staying at the injection site for a long time, The bioinjection material has both advantages. In vivo, autologous tissue enters collagen as a scaffold, and chitosan remains as a frame without being absorbed by the organism for a long period of time, so that the shape at the time of injection is maintained even after regeneration of the autologous tissue in the composite gel. It becomes a biological injection material that can be used. That is, the advantages of collagen and chitosan are synergistically exhibited, self-organized, and can maintain the form at the time of injection, which is an unprecedented bioinjection material.

  According to a second aspect of the present invention, in the bioinjection material according to the first aspect, the composite gel is a chitosan gel that is a chitosan solution gel containing the chitosans and a collagen solution gel that contains the collagen. The bioinjection material is characterized by comprising a mixture of collagen gel or a gelled solution containing the chitosans and the collagen.

  In the second aspect, chitosan gel and collagen gel can be mixed to form a composite gel, and a solution containing chitosans and collagen can be gelled to form a composite gel. It becomes a living body injection material that is safe and can be maintained in the form during injection without being absorbed by the living body for a long period of time.

  According to a third aspect of the present invention, there is provided the biological injection material according to the first or second aspect, wherein the collagen is derived from a marine organism.

  In the third aspect, by using marine organism-derived collagen, there is no problem of infectious diseases and the like, and a bioinjection material that is safe even if injected into a living body.

  According to a fourth aspect of the present invention, there is provided the bioinjection material according to the third aspect, wherein the collagen is extracted from a fish skin, bone, or scale.

  In the fourth aspect, by extracting collagen from fish, there is no problem of infectious diseases and the like, and it becomes a safe living body injectable material even if injected into a living body.

  According to a fifth aspect of the present invention, in the biological injection material according to any one of the first to fourth aspects, the composite gel has a pH of 5.5 to 7.0. is there.

  In the fifth aspect, when the pH is 5.5 to 7.0, the same pH as in the body is obtained, and the bioinjection material is more excellent in biocompatibility.

  According to a sixth aspect of the present invention, in the biological injection material according to any one of the first to fifth aspects, the composite gel contains the chitosans and the collagen in a weight ratio of 90:10 to 20:80. It exists in the bioinjection material characterized by having carried out.

  In such a sixth aspect, the composite gel contains collagen and chitosans in a weight ratio of 90:10 to 20:80, so that it is excellent in biocompatibility and has a slow absorption rate in the living body. It becomes a biological injection material which can hold | maintain a form without being absorbed over a period.

  A seventh aspect of the present invention is the biological injection material according to any one of the first to sixth aspects, wherein the composite gel has a viscosity of 1000 to 20000 mPa · s.

  In the seventh aspect, by setting the viscosity to 1000 to 20000 mPa · s, the bioinjectable material capable of maintaining the form without being absorbed over a long period in which both the fluidity and the form retainability are achieved. Become.

  According to an eighth aspect of the present invention, there is provided the biological injection material according to any one of the first to seventh aspects, wherein the chitosan includes succinylated chitosan.

  In the eighth aspect, by including succinylated chitosan, the bioinjection material can be increased in viscosity and can retain its form without being absorbed for a longer period of time.

  According to a ninth aspect of the present invention, in the bioinjection material according to the eighth aspect, the chitosans are 6-O- (carboxymethyl) chitosan, 6-O- (carboxymethyl) chitin, and N- ( It exists in the bioinjection material characterized by including 3-carboxypropanoyl) -6-O- (carboxymethyl) chitosan.

  Such a ninth aspect comprises 6-O- (carboxymethyl) chitosan, 6-O- (carboxymethyl) chitin, and N- (3-carboxypropanoyl) -6-O- (carboxymethyl) chitosan. Thus, the bioinjection material can retain its form without being absorbed for a longer period of time.

  According to a tenth aspect of the present invention, in the bioinjection material according to the ninth aspect, the chitosans are 50-75 mol% of the 6-O- (carboxymethyl) chitosan, and the 6-O- (carboxyl). A bioinjection material comprising 25 to 50 mol% of methyl) chitin and 0 to 39 mol% of N- (3-carboxypropanoyl) -6-O- (carboxymethyl) chitosan is there.

  In the tenth aspect, by setting each of the ratios as described above, a bioinjectable material that can retain its form without being absorbed for a longer period of time is obtained.

  An eleventh aspect of the present invention is the bioinjection material according to any one of the first to tenth aspects, wherein the composite gel is gelatin, agar, carrageenan, xanthan gum, alginate, locust bean gum, gum arabic, and The bioinjection material is characterized by being gelled with at least one gelling agent selected from the group consisting of anionic polymers.

  In the eleventh aspect, a bioinjection material excellent in biocompatibility can be obtained more easily.

  According to a twelfth aspect of the present invention, there is provided the biological injection material according to any one of the first to eleventh aspects, wherein the collagen contained in the composite gel is crosslinked. .

  In the twelfth aspect, since the collagen is cross-linked, the rate of absorption in the living body is reduced, and the biological injection material that can maintain the form without being absorbed for a longer period of time It becomes.

  A thirteenth aspect of the present invention is a cosmetic / medical bulk material for transplantation into a living body, which comprises a porous material containing chitosans and collagen. It is in.

  In such a thirteenth aspect, by combining collagen having excellent biocompatibility with chitosans that are relatively hardly absorbed in the living body and have the property of easily remaining at the injection site for a long period of time, It is a beauty and medical bulk material that has both advantages. In addition, the use of collagen makes it suitable as a scaffold for cells necessary for tissue regeneration in a living body, and not only can self-organize, but chitosan can be used as a frame without being absorbed by the living body over a long period of time. By remaining, it becomes a bulk material that can maintain the form at the time of transplantation even after the autologous tissue is regenerated in the bulk material. In other words, the advantages of collagen and chitosan are synergistically exhibited, self-organizing, and a bulk material for beauty and medical use that can retain the form at the time of transplantation without being absorbed in vivo. .

  A fourteenth aspect of the present invention is the cosmetic / medical bulk material according to the thirteenth aspect, wherein the porous body contains the collagen in a porous body formed from a chitosan solution containing the chitosans. A bulk material for beauty and medical use, which is impregnated with a collagen solution or a collagen gel that is a gel thereof.

  In the fourteenth aspect, the porous material formed from a chitosan solution containing chitosans is impregnated with a collagen solution containing collagen or collagen gel, which is a gel thereof, so that the chitosans are bulk materials. Collagen that forms the skeleton and is retained in the bulk skeleton facilitates the entry of autologous cells and improves biocompatibility, so that it is easy to adapt to the living body and is not absorbed over a long period of time, so that the form at the time of transplantation It becomes a beauty / medical bulk material that can be held.

  A fifteenth aspect of the present invention is the cosmetic / medical bulk material according to the thirteenth aspect, wherein the porous body is made of a porous material formed from a solution containing the chitosans and the collagen. It exists in the bulk material for beauty and medical treatment characterized by being.

  In the fifteenth aspect, by forming a porous body formed from a solution containing chitosans and collagen, the chitosans form a bulk material skeleton and collagen with excellent biocompatibility is retained in the bulk skeleton. Thus, the bulk material for beauty and medical use can be easily adapted to a living body and can retain the form at the time of transplantation without being absorbed for a long period of time.

  A sixteenth aspect of the present invention is the cosmetic / medical bulk material according to any one of the thirteenth to fifteenth aspects, wherein the collagen contained in the porous body is cross-linked. Located in beauty and medical bulk materials.

  In the sixteenth aspect, since the collagen is cross-linked, it is not absorbed for a longer period of time and can maintain the form at the time of transplantation, and has a mechanical and beauty bulk. Become a material.

  According to a seventeenth aspect of the present invention, there is provided the cosmetic / bulk bulk material according to any one of the thirteenth to sixteenth aspects, wherein the collagen is derived from marine organisms. is there.

  In the seventeenth aspect, by using marine organism-derived collagen, there is no problem such as infectious diseases, and the bulk material for beauty / medical use is safer even if transplanted into the living body.

  An eighteenth aspect of the present invention is the cosmetic / medical bulk material according to the seventeenth aspect, wherein the collagen is extracted from fish skin, bone, or scale. In bulk material.

  In such an eighteenth aspect, the bulk material for beauty / medical use is safer even if transplanted into a living body.

  According to the present invention, by including collagen and chitosans, it is suitable as a cell scaffolding material for safely regenerating tissue even when injected into a living body, and not only can self-organize. It becomes a bioinjection material that is difficult to be absorbed in the living body and can maintain the form at the time of injection at the injection site for a long period of time, or a bulk material for beauty and medical use that can maintain the form at the time of transplantation.

  Hereinafter, embodiments of the present invention will be described in detail.

  The bioinjection material according to the present invention comprises a composite gel containing chitosans and collagen. The composite gel here refers to a gel containing chitosans and collagen in a gel containing a large amount of water and having a certain degree of viscosity. That is, the manufacturing method is not particularly limited as long as it is a gel containing chitosans and collagen. For example, it is obtained by appropriately adding a gelling agent to an aqueous solution containing chitosans and collagen, and a gel obtained by gelling an aqueous solution of chitosans, and a gel obtained by gelling an aqueous solution of collagen Is obtained by mixing. The bioinjection material according to the present invention is in a gel state and has excellent fluidity because of fluidity. Moreover, since it does not diffuse from an injection | pouring site | part at the time of injection | pouring, when it inject | pours in a living body, a form can be hold | maintained. Moreover, even if it inject | pours in any site | part, it can become a suitable form.

  Moreover, the biological injection material concerning this invention consists of a composite gel which has chitosans and collagen as a main component. The main component here means that it is one of the main components exhibiting the function, and does not necessarily indicate that it is the most numerous component. In addition, it is preferable that the composite gel in this invention contains more chitosans than collagen.

  The bioinjection material according to the present invention combines the advantages of both by combining collagen with excellent biocompatibility and chitosans that are difficult to be absorbed in the living body and have the property of easily remaining at the injection site over a long period of time. It has both. Here, collagen is a kind of human dermis, ligament, bone, blood protein, and is the main component of the extracellular matrix, so it is highly biocompatible in vivo, but absorbed in vivo. There is a disadvantage that the speed is high. On the other hand, chitosans are inferior in biocompatibility to collagen, but are absorbed at a very low rate in vivo. By coexisting such chitosans and collagen, the properties of each other are complemented, and the biocompatibility is good due to the properties of collagen at the time of in vivo injection. By remaining unabsorbed, the form at the time of injection can be maintained in vivo for a long period of time. Specifically, in the initial stage of injection into a living body, not only the biocompatibility is improved by collagen, but also the self-organization proceeds into the bio-injecting material by using collagen as a scaffold and self-organization proceeds. . After the collagen is absorbed after a while, the chitosan remains as a frame, and the bioinjection material (into the chitosan) enters and fills with self-organized cells so that it can be injected in vivo for a long period of time. Is maintained. The bioinjection material according to the present invention specifically acts as a scaffold for fibroblasts and vascular endothelium. Since the fibroblasts produce autogenous collagen and extracellular matrix, and blood vessels are regenerated inside the material by the entry of vascular endothelial cells, the bioinjection material according to the present invention is self-organized.

  The composite gel in the present invention is a mixture of a chitosan gel obtained by gelling a chitosan solution containing chitosans with a gelling agent and a collagen gel obtained by gelling a collagen solution containing collagen with a gelling agent as described above. A gelled solution containing chitosans and collagen is preferred. That is, the composite gel is preferably a gel in which both chitosans and collagen are gelled. Moreover, the composite gel may contain other components such as a stabilizer in addition to chitosans and collagen.

  The chitosan solution and the collagen solution here are obtained by, for example, dissolving chitosans or collagen in an aqueous solvent such as water or acetic acid. The chitosan solution may contain other components such as a stabilizer in addition to chitosans, and the collagen solution may contain other components such as a stabilizer in addition to collagen.

  Moreover, the gelatinizer for manufacturing the composite gel in this invention is not specifically limited, A conventionally well-known gelatinizer can be used. For example, gelatin, agar, carrageenan, xanthan gum, alginate, locust bean gum, gum arabic, anionic polymer and the like can be mentioned as the gelling agent for gelling the above-mentioned chitosan solution. The collagen solution can be gelled by adjusting the pH without using a gelling agent.

  The composite gel in the present invention preferably contains 90:10 to 20:80 by weight ratio of chitosans and collagen, more preferably 90:10 to 50:50, particularly preferably 90:10 to 80. : 20. By setting it within this range, the bioinjection material is excellent in biocompatibility and retains an appropriate viscosity, and can maintain its form in vivo. Moreover, the absorption speed | velocity | rate in a biological body becomes slow, and it will not be absorbed over a long period of time. If the proportion of collagen is larger than this range, the viscosity of the bioinjection material tends to be low, and the rate of absorption in the living body may be increased, which is not preferable.

  The viscosity of the composite gel in the present invention is preferably 1000 to 20000 mPa · s. When the viscosity is within this range, it has appropriate fluidity, is easily injected into the living body, and does not diffuse too much in the living body. Moreover, by setting it as the viscosity mentioned above, it can inject | pour, for example with the injection needle of 25-27 gauge used for wrinkle extension. In addition, when the viscosity is higher than this range, it is not preferable because passage of the injection needle, injection into the living body becomes difficult, and fluidity is lost. Further, if the viscosity is lower than this range, the rate of absorption in the living body is increased, which is not preferable.

  The composite gel in the present invention preferably has a pH of 5.5 to 7.0, more preferably pH 6.0 to 6.5. By setting the pH within this range, the pH is about the same as that in the living body, the inflammatory reaction hardly occurs, and the cytotoxicity is low, so that the bioinjection material with high biocompatibility is obtained.

  The collagen in the present invention is preferably derived from marine organisms. Collagen derived from marine organisms, so-called marine collagen, is superior in biocompatibility because it does not cause zoonotic diseases like collagen derived from cattle and pigs. The marine collagen is extracted from, for example, fish skin, bones, scales, shellfish shells, and the like. Here, particularly preferred are salmon, tilapia and the like. Marine collagen is preferably extracted using at least one acid selected from the group consisting of organic acids such as acetic acid and citric acid, or inorganic acids such as hydrochloric acid. Of course, commercially available collagen may be used.

  The collagen contained in the composite gel is preferably crosslinked. The cross-linking of collagen may be performed, for example, after gelation to obtain a collagen gel or after mixing chitosan gel and collagen gel. This is because by cross-linking collagen, the rate of absorption in the living body is reduced, and the living body injection material that is not absorbed for a longer period of time is obtained. The crosslinking method is not particularly limited, and examples thereof include thermal crosslinking, crosslinking by ultraviolet irradiation, crosslinking agent crosslinking, and enzyme crosslinking.

  Furthermore, the heat denaturation temperature of collagen is preferably 35 ° C. or higher, more preferably 40 ° C. or higher. This is because the heat denaturation temperature is equal to or higher than the human body temperature, so that when injected into a living body, there is no fear that the rate of gelatinization due to denaturation by heat and absorption into the living body will be accelerated. Collagen having a heat denaturation temperature of 35 ° C. or higher can be obtained, for example, by increasing the heat denaturation temperature by crosslinking the collagen described above.

  The chitosans in the present invention refer to a product obtained by deacetylating chitin (β-poly-N-acetyl-D-glucosamine) or a derivative thereof, and other than chitosan (β-poly-D-glucosamine) and derivatives thereof. , Which contains unreacted chitin and its derivatives. The chitosans that can be used in the present invention preferably have a chitin deacetylation degree, that is, a chitosan ratio of 50% or more.

  The main components of the chitosans used in the present invention are not particularly limited. For example, chitosan, N-allyl chitosan, N-alkyl chitosan, o-allyl chitosan, o-alkyl chitosan, sulfated chitosan, nitrated chitosan, carboxymethylated Although chitosan etc. are mentioned, chitosan and carboxymethylated chitosan are preferable. By using chitosan having a functional group, it is possible to deliver the bioinjection material at a desired concentration while controlling the time to a specific site in the living body.

  Moreover, it is preferable that chitosan used for this invention contains succinylated chitosan. It is because a chitosan gel can be stably produced by including succinylated chitosan.

  The chitosans in the present invention include, for example, 6-O- (carboxymethyl) chitosan, 6-O- (carboxymethyl) chitin, N- (3-carboxypropanoyl) -6-O- (carboxymethyl) chitosan. Is preferably included. In particular, 6-O- (carboxymethyl) chitosan is 50-75 mol%, 6-O- (carboxymethyl) chitin is 25-50 mol%, N- (3-carboxypropanoyl) -6-O- (carboxy Methyl) chitosan is preferably composed of 0 to 39 mol%. This is because, when the ratio is as described above, an appropriate viscosity is maintained when a chitosan solution containing chitosans is gelled with a gelling agent.

  Here, a chitosan solution containing chitosans composed of 6-O- (carboxymethyl) chitosan, 6-O- (carboxymethyl) chitin, N- (3-carboxypropanoyl) -6-O- (carboxymethyl) chitosan The manufacturing method will be described.

  6-O- (carboxymethyl) chitin is 1 to 10 wt%, preferably 3 to 5 wt% in an alkaline solution containing an alkaline substance such as caustic soda and caustic potash at a temperature of 20 to 100 ° C., preferably 40 to 80 ° C. By reacting for 1 to 24 hours, preferably 3 to 8 hours to deacetylate a part of 6-O- (carboxymethyl) chitin, 6-O- (carboxymethyl) chitosan, 6-O- ( A solution consisting of carboxymethyl) chitin is obtained.

  The solution is neutralized with a mineral acid or an organic acid, and then a succinylating agent such as succinic anhydride is added to react with chitosan.

  Then, the obtained solution is subjected to dialysis membrane treatment, or a precipitate is precipitated with a poor solvent such as methanol or acetone, and the attached salts are separated, whereby 6-O- (carboxymethyl) chitosan, 6-O- A chitosan solution containing chitosans composed of (carboxymethyl) chitin and N- (3-carboxypropanoyl) -6-O- (carboxymethyl) chitosan is obtained.

  Here, an example of the manufacturing method of the composite gel in this invention is demonstrated. A method of mixing a chitosan gel that is a gel of a chitosan solution containing chitosans and a collagen gel that is a gel of a collagen solution containing collagen will be described.

  For example, chitosan gel is obtained by adding water and a gelling agent to a chitosan solution obtained by the method described above or a commercially available chitosan solution and stirring the solution.

  On the other hand, collagen is extracted by using at least one acid selected from the group consisting of organic acids such as acetic acid and citric acid, or inorganic acids such as hydrochloric acid for fish skin, bone, or scales. A collagen solution is prepared by purifying the solution by salting out or the like. And it is made to gelatinize by adding water and a gelatinizer to a collagen solution, or performing pH adjustment, and obtains a collagen gel.

  A composite gel is obtained by mixing this chitosan gel and a collagen gel.

  Moreover, the other example of the manufacturing method of the composite gel in this invention is demonstrated. In addition, the method to gelatinize the solution containing chitosans and collagen is demonstrated.

  For example, a chitosan solution obtained by the above method or a commercially available chitosan solution and a collagen solution obtained by the above method are mixed and stirred. And a composite gel is obtained by adding water and a gelatinizer.

  The bioinjection material according to the present invention can be injected into a living body, and is suitable for use in, for example, wrinkle stretching, breast augmentation, and cheek augmentation.

  The bioinjection material according to the present invention is composed of a composite gel containing chitosans and collagen, and after the collagen is absorbed, the chitosans are not absorbed for a while and remain in the injection site. Can keep long.

  In addition, autologous fibroblasts collected from within a living body may be cultured with the composite gel of the present invention, and used as a bioinjection material according to the present invention. Such a bioinjection material is easily promoted to self-organization in the living body by being injected into the living body, and can regenerate the subcutaneous tissue even in a transplant bed under adverse conditions such as scarring or poor blood circulation. . That is, the biological injection material containing in-vivo autologous fibroblasts can maintain tissue formation semipermanently.

  The bioinjection material according to the present invention can also be used as a carrier for cells other than fibroblasts, adhesion factors, cytokines and the like.

  The bulk material for beauty / medical use according to the present invention is composed of a porous body containing chitosans and collagen. The porous body here has a porosity, that is, a solid having a large number of pores. The bulk material according to the present invention can be freely shaped according to the shape of the tissue defect, and not only maintains a constant shape in the living body, but also consists of a porous body, so that the cells in the living body are centered from the hole. It becomes easy to invade.

  The bulk material according to the present invention is mainly composed of chitosans and collagen. The main component means that it is one of the main components exhibiting the function, and does not necessarily indicate that it is the most numerous component. In addition, it is preferable that the bulk material concerning this invention contains more chitosans than collagen.

  The cosmetic / medical bulk material according to the present invention uses collagen having excellent biocompatibility and chitosans that are difficult to be absorbed by the living body compared to collagen and have properties that are not absorbed by the living body for a long period of time. The bulk material skeleton is mainly formed of chitosans, and the biocompatibility of the bulk material skeleton is improved by collagen.

  Here, collagen is a kind of human dermis, ligament, bone, blood protein, and is the main component of the extracellular matrix, so it is highly biocompatible in vivo, but absorbed in vivo. There is a disadvantage that the speed is high. On the other hand, chitosans are inferior in biocompatibility to collagen, but are absorbed at a very low rate in vivo. In the present invention, the coexistence of such chitosans and collagen complements each other's properties, and the biocompatibility is good due to the properties of collagen at the time of in vivo transplantation. After the collagen is absorbed, the chitosans By remaining unabsorbed for a while, the form at the time of transplantation can be maintained in vivo for a long period of time. Specifically, at the initial stage of transplantation into a living body, not only the biocompatibility is improved by collagen, but also the self-tissue invades into the bulk material using collagen as a scaffold, and self-organization proceeds. Even after the collagen is absorbed after a while, the chitosan remains as a frame and fills the inside of the bulk material (inside the chitosan skeleton) with the self-organized cells so that it can be transplanted in vivo for a long period of time. The form is retained.

  The chitosans and collagen contained in the cosmetic / medical bulk material in the present invention are as described above.

  Moreover, the bulk material for beauty / medical use according to the present invention is suitable as a scaffold material for cells for regenerating tissue in a living body. Specifically, it can be used as a scaffold for fibroblasts and vascular endothelial cells in vivo. Since these fibroblasts produce self-collagen and extracellular matrix, and vascular endothelial cells form blood vessels, the bulk material for beauty and medical treatment according to the present invention is self-organized. Specifically, collagen adheres fibroblasts and vascular endothelial cells, promotes cell proliferation, and fibroblasts that have entered the bulk material secrete autologous collagen and extracellular matrix, and vascular endothelium. Cells develop blood vessels. For this reason, self-organization progresses in the bulk material according to the present invention. Chitosans are not absorbed for a while after the collagen is absorbed, retain the shape at the time of transplantation, and work as a skeleton of the regenerated tissue. As described above, the bulk material according to the present invention is not absorbed over a long period of time, and can retain the form formed in accordance with the tissue defect portion.

  The porous body that can be used as a cosmetic / medical bulk material according to the present invention is, for example, a collagen solution containing collagen or a gel thereof, which is formed by freeze-drying a chitosan solution containing chitosans. It can be obtained by impregnating the gel or freeze-drying a solution containing chitosans and collagen. The means for obtaining a porous body by making it porous is not limited to freeze-drying.

  Moreover, the porous body used as the beauty / medical bulk material according to the present invention may contain, for example, additives for improving properties as a bulk material in addition to chitosans and collagen. Furthermore, the porous body that can be used as the cosmetic / medical bulk material according to the present invention is not limited to the above-described porous material, and includes, for example, a collagen obtained by freeze-drying a chitosan solution and collagen and chitosans. A solution or gel thereof may be contained, or a collagen solution or gel thereof may be further impregnated into a porous material obtained by freeze-drying a solution containing collagen and chitosans.

  A porous material formed by lyophilizing a solution containing chitosans and collagen can be obtained by lyophilizing the chitosan solution and then impregnating the collagen solution and lyophilizing it. It may be impregnated and freeze-dried, or may be freeze-dried after mixing the collagen solution and the chitosan solution.

  The porous body can also be obtained by lyophilization after adding a collagen solution containing collagen or chitosan fiber containing chitosan to a collagen porous material and homogenizing.

  The porous material in the present invention is preferably a porous material having a high porosity and a relatively large pore diameter so that autologous cells can easily enter. The porous material preferably has a pore diameter of about 50 μm to 400 μm, a porosity of about 50 vol% to about 80 vol%, and a pore-to-pore communication hole diameter of about 30 μm to 50 μm. This is because the cells easily enter the bulk material, so that soft tissues and the like in the living body are easily formed inside, that is, self-organization easily proceeds.

  In addition, it is preferable that the collagen contained in the porous body in the present invention is crosslinked by thermal crosslinking, crosslinking by ultraviolet irradiation, crosslinking agent crosslinking, enzyme crosslinking, or the like. This is because collagen is cross-linked to reduce the rate of absorption in vivo. Moreover, it is because the mechanical strength of a porous body improves by bridge | crosslinking collagen.

  In addition, the porous body in the present invention is not limited to the above-described one, and for example, a porous body including a chitosan-containing porous body and a collagen-containing porous body may be laminated (laminated). At this time, it is preferable to mold so that the porous material containing chitosans is sandwiched between the porous materials containing collagen. Further, by laminating (laminating) a plurality of layers, the thickness and shape of the porous body can be freely controlled and self-organized into an arbitrary shape.

  The bulk material for beauty / medical use of the present invention is used by being embedded in a desired part of a living body, and is used by forming the porous body produced by the above-described various methods into a desired shape. .

  An example of the manufacturing method of the bulk material concerning this invention is demonstrated. Here, the bulk material which consists of a porous body formed by freeze-drying the solution containing chitosans and collagen is demonstrated. However, the manufacturing method of a bulk material is not limited to this.

  First, a chitosan solution is obtained by dissolving chitosans obtained by treating a crustacean shell or the like with hydrochloric acid in an organic acid such as acetic acid or lactic acid, or an inorganic acid such as hydrochloric acid. Of course, a commercially available chitosan solution may be used.

  Next, after lyophilizing the chitosan solution, an alkaline solution such as NaOH is added and stirred and washed with water. As a result, the chitosans are insolubilized by neutralizing the acid.

  Thereafter, this is freeze-dried again to obtain a porous material composed of chitosans.

  Next, a collagen solution is impregnated into a porous material made of chitosans. For example, collagen is extracted from fish skin, bones, scales, etc. with acetic acid, hydrochloric acid, citric acid or the like, and purified by salting out to obtain a collagen solution. Of course, a commercially available collagen solution may be used.

  Finally, the porous material impregnated with the collagen solution is lyophilized and heated to thermally crosslink the collagen to obtain a bulk material.

  The bulk material according to the present invention can be made into a thick sponge-like bulk material by a freeze-drying method or the like in the manufacturing process. That is, it is possible to obtain a bulk material having high porosity, high pore diameter, and good communication that satisfies the above-mentioned criteria. For this reason, the bulk material according to the present invention is easy for cells to enter into the center.

  Conventional wound dressings were mostly artificial skin intended for epithelialization of shallow skin defects, but the bulk material according to the present invention can be used not only for breast augmentation, cheek augmentation, Transplantation into congenital / acquired tissue defects, ie, large defects in soft tissue can be compensated. Moreover, if it is used as a carrier for various cells, nutrient factors, adhesion factors, and cytokines, it is effective not only for soft tissue but also for reconstruction of hard tissue.

  Hereinafter, the present invention will be described based on examples. However, the present invention is not limited to this.

<Bioinjection material 1>
(Example 1)
28 g of 6-O- (carboxymethyl) chitin was dissolved in 1400 g of a 6 wt% NaOH solution and stirred at 70 ° C. for 6 hours to deacetylate the chitin. After cooling this solution to room temperature, the pH was adjusted to 6.5, 1.0 g of succinic anhydride was added, and the mixture was stirred at 50 ° C. for 2 hours to succinylate part of the chitosan contained in the solution. Thereafter, the attached salts are desalted and purified with a dialysis membrane, so that N- (3-carboxypropanoyl) -6-O- (carboxymethyl) chitosan is 3 mol% and 6-O- (carboxymethyl) chitosan is A chitosan solution comprising 62 mol% and 35 mol% 6-O- (carboxymethyl) chitin was obtained.

  To 16 g of 1 wt% chitosan solution, 9 g of 0.5 wt% carboxyvinyl polymer liquid as a gelling agent and 65 g of water were added and stirred to obtain a chitosan gel.

  Collagen was extracted from salmon skin, bones and scales, purified by salting out, freeze-dried, dissolved in dilute hydrochloric acid and dialyzed to change the pH to obtain 10 g of 0.5 wt% collagen gel.

  Collagen gel was added to this chitosan gel and stirred to obtain the bioinjection material of Example 1.

(Example 2)
Except that 9 g of 0.5 wt% carboxyvinyl polymer solution as a gelling agent and 3 g of 1,3-butylene glycol as a stabilizer and 62 g of water were added to 16 g of 1 wt% chitosan solution and stirred to obtain chitosan gel. In the same manner as in Example 1, a biological injection material of Example 2 was obtained.

(Example 3)
The same procedure as in Example 1 was repeated except that 14 g of 0.5 wt% carboxyvinyl polymer solution as a gelling agent and 52 g of water were added to 24 g of 1 wt% chitosan solution and stirred to obtain chitosan gel. A bioinjection material was obtained.

Example 4
24 g of 1 wt% chitosan solution was mixed with 14 g of 0.5 wt% carboxyvinyl polymer solution as a gelling agent, 5 g of 1,3-butylene glycol as a stabilizer and 47 g of water, and stirred to obtain chitosan gel. In the same manner as in Example 1, a biological injection material of Example 4 was obtained.

  Table 1 shows the composition and characteristics of the biomaterials of Examples 1 to 4.

  As shown in Table 1, the biological injection materials of Examples 1 to 4 had a pH of 6.1 to 6.3 and a viscosity of 1300 mPa · s to 15500 mPa · s.

  In addition, the photograph at the time of dripping the biological injection material of Example 4 to a plate is shown in FIG. FIG. 1A is a photograph when dropped on a horizontal plate, and FIG. 1B is a photograph when dropped on an inclined plate.

  As shown in FIG. 1, the bioinjection material of Example 4 has a viscosity that does not flow down even when the plate is tilted. Thus, it is considered that the bioinjection material of the present invention does not diffuse from the injection site when injected into the living body.

<Bioinjection material 2>
(1) Chitosan gel 3 mol% of N- (3-carboxypropanoyl) -6-O- (carboxymethyl) chitosan and 62 mol% of 6-O- (carboxymethyl) chitosan were obtained in the same manner as in Example 1. A chitosan solution consisting of 35 mol% of 6-O- (carboxymethyl) chitin was obtained. To this chitosan solution, 0.5 wt% carboxyvinyl polymer solution as a gelling agent, 1,3-butylene glycol as a stabilizer and water were added and stirred to obtain a chitosan gel.

(2) Collagen gel Collagen was extracted from tilapia scales, dissolved in water, and then thermally crosslinked to obtain a collagen solution having a denaturation temperature of 35 to 37 ° C. The collagen solution was purified by salting out, freeze-dried, dissolved in dilute hydrochloric acid, and dialyzed to change the pH to obtain a collagen gel.

(Example 5)
The bioinjection material of Example 5 was obtained by mixing chitosan gel and collagen gel so that the weight ratio of chitosans and collagen was 60:40.

(Example 6)
The bioinjection material of Example 6 was obtained by mixing chitosan gel and collagen gel so that the weight ratio of chitosans and collagen was 70:30.

(Example 7)
The bioinjection material of Example 7 was obtained by mixing chitosan gel and collagen gel so that the weight ratio of chitosans and collagen was 80:20.

(Example 8)
The bioinjection material of Example 8 was obtained by mixing chitosan gel and collagen gel so that the weight ratio of chitosans and collagen was 90:10.

(Comparative Example 1)
Chitosan gel was used as the bioinjection material of Comparative Example 1.

(Comparative Example 2)
Collagen gel was used as the bioinjection material of Comparative Example 2.

  Table 2 shows the compositions and characteristics of the biomaterials of Examples 5 to 8 and Comparative Examples 1 and 2.

(Test Example 1)
1 ml of the bioinjection materials of Examples 5 to 8 and Comparative Examples 1 and 2 were injected subcutaneously into the back of 24 Wistar rats, three by three. In addition, a different bioinjection material was injected into each of the three locations.

  Two animals were collected from each group 3 days, 1 week and 2 weeks after the operation. Two samples from each group were divided into two halves, one with HE-stained specimens (LM) and the other stored in fixative. Observation was performed with a TEM (transmission electron microscope) as necessary.

  2 to 5 show the results of observation of rat subcutaneous tissue 3 days and 2 weeks after the operation in FIGS. 2 to 5, and the results of HE sample observation of the subcutaneous tissue 1 week after surgery of the rat injected with the bioinjection material of Example 6. As shown in FIG. A photograph of the rat subcutaneous tissue 2 weeks after the operation is shown in FIG.

(Summary of results)
As shown in FIG. 7, all the subcutaneous tissues of rats injected with the bioinjectors of Examples 5 to 8 and Comparative Examples 1 to 2 had mild gross inflammatory findings.

  As is clear from FIG. 2, the bioinjection material of Comparative Example 2 made of collagen gel was almost absorbed on the third day after the operation. On the other hand, the living body injecting materials of Examples 5 to 8 and Comparative Example 1 were not completely absorbed by the living body and maintained the form at the time of injection.

  As shown in FIG. 3, the small circular cell infiltration of the biological injection materials of Examples 5 to 8 was enhanced immediately after the injection, but the cell infiltration into the biological injection materials of Examples 7 and 8 having a high chitosan ratio was mild. was. Cell infiltration into the bioinjection material of Examples 5 and 6 having a high collagen ratio was relatively strong. This is because a large number of phagocytic cells are mobilized to engulf collagen.

  In contrast, the bioinjection material of Comparative Example 1 consisting only of chitosan gel had infiltrated small circular cells into the gel. This is because chitosan induces an inflammatory reaction.

  In the subcutaneous tissue 1 week after surgery of the rat injected with the bioinjection material of Example 6 (FIG. 6), small circular cells infiltrate from the periphery toward the inside of the gel, which is vigorous angiogenesis (V in the figure). Was accompanied. Almost no film formation was observed.

  As shown in FIGS. 4 and 5, the bioinjection material of Comparative Example 1 could not be confirmed 2 weeks after the operation. On the other hand, the biological injection material of Examples 5-8 was maintaining the form at the time of injection | pouring.

  From the above, it can be seen that a bioinjection material composed of a gel composed of chitosans and collagen is safer and less easily absorbed by the living body than injecting a single gel into the living body and remains at the injection site for a long period of time. It was. Moreover, it turned out that the bioinjection material which contains chitosans and collagen 80: 20-90: 10 by weight ratio is especially excellent in biocompatibility.

<Bulk material>
Example 9
100 g of 1.5 wt% chitosan acetic acid solution (manufactured by Hokkaido Soda Co., Ltd.) was lyophilized at −10 ° C. for 48 hours to obtain 2.15 g of white plate-like porous material. This was immersed in 4.2 wt% sodium hydroxide (manufactured by Wako Pure Chemical Industries, Ltd.) for 1 hour at room temperature, and then washed with distilled water. This was freeze-dried at −10 ° C. for 48 hours to obtain a porous material composed of chitosans.

  Next, a porous material composed of chitosans was immersed in 100 g of a 0.5 wt% collagen-derived collagen solution (manufactured by Ihara Suisan Co., Ltd.) for 1 hour at room temperature to allow the collagen solution to permeate and absorb. Furthermore, after freeze-drying this at -10 degreeC for 48 hours, it heated at 140 degreeC for 16 hours with the vacuum dryer, and the bulk material of Example 9 was obtained by performing thermal crosslinking. A flow chart of the bulk material manufacturing method is shown in FIG. 8, and a photograph is shown in FIG.

(Example 10)
A 0.5 wt% koji-derived collagen solution 100 g (manufactured by Ihara Suisan Co., Ltd.) was freeze-dried at −10 ° C. for 48 hours to obtain a porous white plate-like collagen. The resulting porous collagen was heated and crosslinked at 140 ° C. for 16 hours in a vacuum dryer. The porous layer made of collagen was immersed in 100 g of a 5.0 wt% chitosan acetic acid solution (manufactured by Hokkaido Soda Co., Ltd.) for 2 hours at room temperature, and the chitosan acetic acid solution was permeated and absorbed into the porous layer made of collagen. This was freeze-dried at −10 ° C. for 48 hours, immersed in 4 wt% sodium hydroxide (manufactured by Wako Pure Chemical Industries, Ltd.) for 1 hour at room temperature, and washed with distilled water. The washed composite was lyophilized at −10 ° C. for 48 hours to obtain the bulk material of Example 10. A flow chart of the bulk material manufacturing method is shown in FIG. 8, and a photograph is shown in FIG.

(Example 11)
A 0.5 wt% koji-derived collagen solution 128 g (manufactured by Ihara Suisan Co., Ltd.) and a 1.5 wt% chitosan acetic acid solution 100 g (manufactured by Ihara Suisan Co., Ltd.) were stirred at room temperature to prepare a chitosan-collagen mixed solution. This solution was freeze-dried at −10 ° C. for 48 hours to obtain chitosan-collagen porous material. The obtained chitosan-collagen porous material was heated at 140 ° C. for 16 hours in a vacuum dryer, thermally crosslinked, and immersed in 4.2 wt% caustic soda (Wako Pure Chemical Industries, Ltd.) at room temperature for 1 hour. Then, it was washed with distilled water. The washed porous material was freeze-dried at −10 ° C. for 48 hours to obtain a bulk material of Example 11. A flow chart of the bulk material manufacturing method is shown in FIG. 8, and a photograph is shown in FIG.

(Example 12)
North chitosan MC-2W (manufactured by Hokkaido Soda Co., Ltd.) was dissolved in acetic acid to prepare a 0.5 wt% chitosan acetic acid solution. The chitosan acetic acid solution and 0.5% wt% collagen solution were mixed so that the ratio was 7: 3, and 100 ml of this mixed solution was placed in a 5 × 3.5 cm tray. This was freeze-dried to obtain a porous material. Water-soluble carbodiimide as a cross-linking agent was dissolved in 4M NaCl, and the porous material was subjected to a cross-linking reaction at 4 ° C. for 48 hours. Thereafter, the porous material was washed with distilled water. The washed porous material was freeze-dried to obtain a bulk material of Example 12. A flowchart of the bulk material manufacturing method is shown in FIG.

(Example 13)
A bulk material of Example 13 was obtained in the same manner as Example 12 except that the ratio of the chitosan solution to the collagen solution was 5: 5.

(Example 14)
A bulk material of Example 14 was obtained in the same manner as Example 12 except that the ratio of the chitosan solution and the collagen solution was 3: 7.

(Example 15)
A chitosan trifluoroacetic acid solution having a degree of deacetylation of 93% was placed in a syringe and connected to a 22G needle, the liquid feed rate of the infusion pump was 2 ml / hour, and an electrode plate made of aluminum (5.5 cm × 5.5 cm) A chitosan fiber was obtained on the electrode plate by applying a voltage of 27 kv between the needle and a high-voltage DC voltage power source and ejecting a trifluoroacetic acid solution onto the electrode plate. The chitosan fiber peeled from the electrode plate was immersed in 28% by weight of ammonia water for 1 hour at room temperature, and then distilled water was continuously supplied for 2 hours at room temperature to obtain a chitosan fiber.

  To the chitosan fiber was added collagen porous material that had been thermally crosslinked by heating at 140 ° C. for 16 hours in a vacuum dryer, and homogenized to obtain a slurry liquid. This was freeze-dried at −10 ° C. for 48 hours with a freeze dryer to obtain a bulk material of Example 15. A flowchart of the bulk material manufacturing method is shown in FIG.

(Example 16)
A 0.5 wt% koji-derived collagen solution is added to the chitosan fiber obtained in Example 15 and homogenized to obtain a slurry solution. This was freeze-dried at −10 ° C. for 48 hours with a freeze dryer and then heated at 140 ° C. for 16 hours with a vacuum dryer to carry out thermal crosslinking to obtain a bulk material of Example 16. A flow chart of the bulk material manufacturing method is shown in FIG. 8, and a photograph is shown in FIG.

(Test Example 2)
The thickness and density of the bulk materials of Examples 9 to 11 were measured. The results are shown in Table 3.

  As shown in Table 3 and FIGS. 9 to 11, the bulk materials of Examples 9 to 11 were all sponge-like and low density. As a result, the bulk material of the present invention comprising a porous material containing chitosans and collagen can be made to have a low density that is easy for cells in the body to invade and has a thickness. I found that it was possible. By appropriately adjusting the conditions, such as changing the proportion of collagen, the absorption rate in the living body can be changed.

It is a photograph at the time of dripping the biological injection material of Example 4 on a plate. It is an enlarged photograph of the subcutaneous tissue of a rat 3 days after surgery. It is the photograph which expanded further the enlarged photograph of FIG. It is an enlarged photograph of the subcutaneous tissue of a rat 2 weeks after the operation. It is the photograph which expanded further the enlarged photograph of FIG. It is an enlarged photograph of the subcutaneous tissue of a rat 1 week after inject | pouring the biological injection material of Example 6. FIG. It is a photograph of the subcutaneous tissue of a rat 2 weeks after the operation. It is a flowchart which shows the manufacturing method of the bulk material concerning this invention. 10 is a photograph of the bulk material of Example 9. 10 is a photograph of a bulk material of Example 10. 10 is a photograph of the bulk material of Example 11. It is a photograph of the bulk material of Example 16.

Claims (18)

A bio-injection material comprising a composite gel containing chitosans and collagen. The bioinjection material according to claim 1, wherein the composite gel is a mixture of a chitosan gel that is a chitosan solution gel containing the chitosans and a collagen gel that is a collagen solution gel containing the collagen, or A bioinjection material comprising a gelled solution containing the chitosans and the collagen. The biological injection material according to claim 1 or 2, wherein the collagen is derived from marine organisms. 4. The bioinjection material according to claim 3, wherein the collagen is extracted from fish skin, bones, or scales. The biological injection material according to any one of claims 1 to 4, wherein the composite gel has a pH of 5.5 to 7.0. The bioinjection material according to any one of claims 1 to 5, wherein the composite gel contains the chitosans and the collagen in a weight ratio of 90:10 to 20:80. Wood. The biological injection material according to any one of claims 1 to 6, wherein the composite gel has a viscosity of 1000 to 20000 mPa · s. The biological injection material according to any one of claims 1 to 7, wherein the chitosan contains succinylated chitosan. The bioinjection material according to claim 8, wherein the chitosans are 6-O- (carboxymethyl) chitosan, 6-O- (carboxymethyl) chitin, and N- (3-carboxypropanoyl) -6-O. A bioinjection material comprising-(carboxymethyl) chitosan. The bioinjection material according to claim 9, wherein the chitosan is 50 to 75 mol% of the 6-O- (carboxymethyl) chitosan, 25 to 50 mol% of the 6-O- (carboxymethyl) chitin, A bioinjection material, wherein the N- (3-carboxypropanoyl) -6-O- (carboxymethyl) chitosan comprises 0 to 39 mol%. The bioinjection material according to any one of claims 1 to 10, wherein the composite gel is selected from the group consisting of gelatin, agar, carrageenan, xanthan gum, alginate, locust bean gum, gum arabic, and an anionic polymer. A biological injection material characterized in that it is gelled with at least one gelling agent. The biological injection material according to any one of claims 1 to 11, wherein the collagen contained in the composite gel is cross-linked. A bulk material for beauty / medical use for transplanting into a living body, comprising a porous material containing chitosans and collagen. 14. The cosmetic / medical bulk material according to claim 13, wherein the porous body is impregnated with a collagen solution containing the collagen or a collagen gel that is a gel thereof in a porous material formed from a chitosan solution containing the chitosans. A bulk material for beauty / medical use characterized by The cosmetic / medical bulk material according to claim 13, wherein the porous body is a porous material formed from a solution containing the chitosans and the collagen. . The bulk material for beauty / medical use according to any one of claims 13 to 15, wherein the collagen contained in the porous body is cross-linked. The bulk material for beauty / medical use according to any one of claims 13 to 16, wherein the collagen is derived from marine organisms. The bulk material for beauty and medical treatment according to claim 17, wherein the collagen is extracted from fish skin, bone, or scale.

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