KR20030097156A - Tissue Engineered Natural/Synthetic Hybrid Scaffolds and its Manufactory Methods - Google Patents

Abstract

PURPOSE: Provided are a porous/biodegradable carrier which is produced by combining a tissue under mucous membrane of small intestine or a bone powder of mammals with synthetic polymer, and a method for producing the same. CONSTITUTION: The carrier is produced by combining a biomaterial such as tissue under mucous membrane of small intestine or bone powder of mammals with synthetic polymer. The mammals are selected from cattle, pig, rabbit, mouse, and so on. The synthetic polymer is at least one of chitin, chitosan, alginic acid, polylactic acid(PLA), polyglycolic acid(PGA), lactic acid-glycolic acid copolymer(PLGA), polyhydroxybutyric acid, polycaprolactone, polyanhydride, and polyalkylcyano acrylate. The carrier has both advantages of biodegradable synthetic polymer carrier and natural material.

Description

Tissue Engineered Natural / Synthetic Hybrid Scaffolds and its Manufactory Methods}

The present invention relates to a natural / synthetic biodegradable hybrid carrier for tissue engineering and a method of manufacturing the same, and more particularly, using a biological tissue such as small intestinal submucosa or bone meal as a material, superior wettability to conventional synthetic biodegradable polymer carriers. Biodegradable synthetic polymer carriers and natural products not only have very favorable conditions for sowing cells but also promote tissue regeneration by sustained release of bioactive substances such as growth factors or cytokines contained in small intestinal submucosa or bone meal. The present invention relates to a natural / synthetic biodegradable hybrid carrier for tissue engineering, a method for preparing the same, and an artificial organ using the carrier, having all the advantages of the material.

Despite the rapid growth of modern, sophisticated and sophisticated medical engineering technologies, diseases that damage human organs and tissues are treated with various accidents and the advancement of the level of health care. The costs are high, and they are emerging throughout the society at relatively high frequency and with serious problems. When some organs or parts of the body are damaged by diseases or accidents, artificial organs are used to treat patients' diseases. The development history of these artificial organs, which have been around 100 years since Lane was used to fix bone fractures around 1890, can be divided into four generations.

The first generation was a very early transplant prior to 1940 that supported or prostheted some of the primitive metals and ceramics in the human body. These studies or procedures began with a basic curiosity rather than practicality or effects without much distinction between disciplines such as medicine, chemistry and materials.

The second generation begins with doctors replacing a part of a broken or damaged organ with a generic chemical material devised by researchers, accidentally and unexpectedly without an exact definition of abnormality, ie biocompatibility with tissues that come into contact with the human body. It was a time when it was applied. For example, Chanley uses polymethyl methacrylate bone cement as artificial hip joint, and Boris Lee uses nylon and polyacrylonitrile-based copolymer as artificial blood vessel in Korean war, plasticizing with diethylhexyl phthalate. This is an example of using vinyl chloride as a blood bag and using polyvinylpyrrolidone as an artificial blood during the Vietnam War. These examples have been widely used since the late 1950, when the operation was performed in earnest, with numerous clinical successes.

The third generation is the body and the "bioactive" state, i.e. very precisely designed and applied polymers, ceramics and metal materials, rather than the "inactive" state without any interaction with the living body highlighted in the first and second generations described above. Is a developmental generation of biomaterials that stimulate surrounding tissue cells to make transplantation faster and more effective. Typical examples include artificial hip joints coated with hydroxyapatite, artificial blood vessels coated with algin and collagen, and the like.

The fourth generation is the development of mixed artificial organs using tissue engineering, that is, tissue cells extracted from the human body and synthetic materials are used simultaneously. They focus on improving and restoring damaged tissues, rather than completely replacing organs of the body with reoperations. The fundamental reason for the development of artificial organs with the fourth generation of tissue engineering is that the general limitations of general synthetic polymers, namely, biocompatibility and the biofunctionality of damaged organs or tissues of a certain part, are eventually lacked. . Accordingly, the company is pursuing a hybrid that mimics biological elements by binding, immobilizing, and culturing cells or proteins, which are responsible for specific functions of organs, to non-degradable polymer materials to further enhance and functionalize desired tissues or organs. .

As a representative example of such a histological study, the artificial cartilage is schematically described as follows. First, a specific organ shape in the human body is sculpted with gypsum or the like, and then a long-term mold is manufactured using copper plate and silicon. A carrier to be supported and grown by chondrocytes in the mold of the organ form must be prepared. As such a carrier, microporous and biodegradable polymers are usually used. Recently, biodegradable polymers as synthetic polymers are used as ester homopolymers and copolymers of lactide and glycolide, and their biodegradation mechanism is due to hydrolysis.

Polylactide (PLA) is used to manufacture bone fixation plates and screws to fix broken bones, and polyglycolide (PGA) is widely used as an absorbent suture. Although the biodegradation period of these homopolymers is somewhat different depending on the molecular weight, it is usually 1 year or more, whereas their copolymer, lactide-glycolide copolymer (PLGA), undergoes biodegradation over several weeks to several weeks depending on the molecular weight. As a method for producing a microporous and biodegradable carrier (for example, a sponge-like structure) using a PLGA biodegradable polymer, it is mixed with a single crystal salt of a predetermined size in a solution in which PLGA is dissolved, and the salt is dried in water. Salt extraction method to dissolve, expansion of PLGA using carbon dioxide, non-woven fabric of PGA fiber to make PGA mesh, evaporation of solvent in PLGA solution, solvent in PLGA solution Phase separation methods, such as immersion in phases, are applied. Then, cartilage cells isolated and mass cultured from rabbit ears are harvested through a series of operations and seeded in ear and nose biodegradable carriers. And, when transplanted into the laboratory animals, the biodegradable polymer is naturally absorbed in the animal body and finally only cartilage is left to replace the role of the lost organs. At this time, hepatocytes, small intestine cells, urinary tract cells, vascular endothelial cells, bone marrow cells, nerve cells, etc. are applied as tissue cells to be separated and seeded, and thus, space, artificial field, artificial urinary tract, artificial blood vessel, artificial bone marrow, artificial It can be applied to nerves and other organs.

On the other hand, the present inventors have proposed a porous, biodegradable artificial organs and a method for producing the same by the emulsion freeze drying method consisting of a polymer solution and water to solve the other disadvantages (Korean Patent No. 201,874). However, the manufacturing method according to Korean Patent No. 201,874 has a disadvantage in that the surface properties of the synthetic biodegradable polymer used are hydrophobic so that the culture solution and cells do not penetrate and grow on the biodegradable polymer carrier. In addition, when the biodegradable polymer carrier contains cytokines of various growth factors, various properties such as angiogenesis and neuronal cell formation can be remarkably increased, but bioactive substances are denatured and water is produced during the manufacturing process. And other solvents pre-flowed out and lost their original function. Moreover, the growth factor or cytokine price is very expensive, so if you use it, you have to have a lot of economic burden.

In addition, in the case of the synthetic polymer carrier for tissue engineering, since most of the surface properties of the synthetic polymer, which is one of the most problematic problems during cell seeding, have a hydrophobic surface, it acts as a factor that prevents the cell suspension from penetrating into the carrier. This means that the cells are dispersed evenly in the carrier, making it difficult to adhere and grow. In order to solve this problem, a method of pre-wetting synthetic polymer carrier in ethanol and then substituting with water [A. G. Mikos, M. D. Lyman, L. E. Freed, and R. Langer, Biomaterials, 15, 55 (1994)] and a surface modified by treating the carrier with NaOH solution [J. Gao, L. Niklason, and R. Langer, J. Biomed. Mater. Res., 42, 417 (1998)], a method for treating with a mixed solution of chloric acid [S. J. Lee, G. Khang, Y. M. Lee, and H. B. Lee, J. Biomater. Sci. Polymer Edn., 13, 197 (2002)] and many other physicochemical surface modification methods have been used, but this has been pointed out as a problem of reducing the physical properties of the carrier or the toxicity of the remaining solvent. Due to these problems, materials such as collagen, chitosan, and chitin, which are natural materials, have been used, but this also has problems such as not only complicated separation process but also poor processability and weak physical properties compared to synthetic polymers.

Meanwhile, in the case of artificial organs of No. 2000-13700, which have been patented by the present inventors, porous and biodegradable polymers together with physiologically active substances were prepared by emulsification freeze drying method. Due to its high price and very low stability to solvents, it is very inefficient and its precise mechanism for its function is not proven, and because it contains very small amount, the problem of synthetic polymer itself is not solved. It is necessary to manufacture a porous, biodegradable artificial organ with excellent processability.

Accordingly, the present inventors have studied to solve the above problems, and as a result, to solve the problem that most of the polymer material has a hydrophobic problem, and to develop a material that has a beneficial effect on the growth, differentiation and migration of cells. The mammalian small intestinal submucosa and bone meal are mostly composed of collagen and contain excessive amounts of physiologically active substances, which facilitates tissue regeneration. Thus, the present invention has been developed by complexing them with biodegradable polymers to develop porous and biodegradable carriers for tissue engineering. It was completed.

Accordingly, an object of the present invention is to provide a porous biodegradable carrier prepared by complexing a mammalian small intestinal submucosa or bone powder with a synthetic polymer and a method for producing the same.

In addition, another object is to provide an artificial organ prepared using the porous biodegradable carrier.

1 is a scanning electron micrograph showing the surface (a) and cross-section (b) of the small intestinal submucosa / synthetic polymer complexed porous and biodegradable carrier according to the present invention,

2 is a scanning electron micrograph showing a surface (a) and a cross-section (b) of a bone powder / synthetic polymer composite porous / biodegradable artificial organ according to the present invention;

3 is a test for wetting the existing synthetic polymer carrier and the small intestinal submucosa / synthetic polymer composite porous and biodegradable carrier according to the present invention,

Figure 4 shows the water absorption of the conventional composite polymer carrier and the small intestinal submucosa / synthetic polymer composite porous, biodegradable carrier according to the present invention,

Figure 5 shows the cell adhesion test according to the content of the existing synthetic polymer carrier and small intestinal submucosa according to the present invention,

Figure 6 is a picture (a) of the fibroblasts cultured in the basic culture, a picture (b) of the fibroblasts cultured with the culture medium to which the extract of the small intestinal submucosa is added,

7 is a photograph of 4 weeks after the small intestinal submucosal tissue / synthetic polymer in which the cells are seeded, and the composite porous / biodegradable organ.

Fig. 8 is a photograph (a) after cell seeding of a conventional synthetic polymer and a photograph (b) of a small intestinal submucosa / synthetic polymer-composite porous / biodegradable artificial organ,

9 is a safranin-O staining picture of histological artificial cartilage prepared with a carrier according to the present invention,

10 is a Bon-Kusa stained picture of histological artificial bones prepared with a carrier according to the present invention,

Figure 11 is a histological staining picture of histological artificial bones made of bone / synthetic polymer carrier of the mammal according to the present invention.

The present invention is characterized by a porous and biodegradable carrier prepared by complexing a small intestinal submucosa or bone powder of a mammal with a biodegradable synthetic polymer and a method for producing the same.

It also includes an artificial organ prepared using the porous biodegradable carrier.

Referring to the present invention in more detail as follows.

The present invention has superior wettability by complexing natural materials than biodegradable synthetic polymer carriers, and has very favorable conditions for sowing of cells, and most of materials are made of proteins such as collagen, so that the adhesion and growth of cells are better. Porous, biodegradable carriers incorporating biodegradable synthetic polymers into biodegradable synthetic polymers that are superior to tissue regeneration due to sustained release of physiologically active substances such as growth factors or cytokines contained in the small intestine submucosa or bone meal And an artificial organ using the carrier.

The small intestinal submucosa used in the present invention is a collagen as a main component, in addition to transforming growth factor (TGF), insulin-like growth factor (IGF), and fibroblast growth factor (acidic and basic fibroblast). growth factor, aFGF and bFGF), vascular endothelial growth factor (VEGF), nerve growth factor (NGF), and glycosaminoglycans (hyronic acid, chondroitin sulfate) , Heparin and the like), fibronectin and the like. These small intestinal submucosa can absorb more than 90% of moisture, and since there are few cells, the immune response is very small and biodegradable. It can be said.

In addition, the bone component of the mammal used in the present invention is composed of 30 to 35% of organic components and 65 to 70% of inorganic components, the organic components are mostly fibrous protein and collagen type I and bone morphogenic protein (bone morphogenic) protein, BMP), and the inorganic component is mainly composed of calcium phosphate. Bone powder used in the present invention was used in the state in which the inorganic component of the bone component is removed.

Biodegradable synthetic polymers used in tissue engineering include polyglycolide (PGA), polylactide (PLA), lactic acid-glycolic acid copolymer (poly (lactide- co- glycolide), PLGA), and polycaprolactone ( Poly (ε-caprolactone), PCL), etc. are used. The advantages of these synthetic polymers are that they have excellent processability and can control the biodegradation period. However, synthetic polymers have a number of modification methods have been preceded by the disadvantage of affinity with cells.

Materials used as natural polymers include collagen (types I, II, III, and IV), fibrin, chitosan, chitin, and the like, and since they are natural materials, they have relatively high affinity with cells. However, there is a problem in that the separation process in the tissue is very complex, poor processability and weak physical properties, there is a problem in the long-term application that requires a lot of force.

The natural / synthetic polymer hybrid carrier proposed in the present invention has been developed to maximize the advantages by reinforcing the disadvantages of the synthetic polymer and natural polymer. There is no series of complex separation processes and complexed with synthetic polymers to improve cell affinity, processability, high bioactivity, and low cost.

Mammalian bone flour or submucosal tissue was isolated and mechanically pulverized into a solution containing dissolved bioactive substances, and fibroblasts were cultured using the isolated extracts. It was confirmed that there are factors that have a major influence on cell adhesion and growth.

By using such small intestinal submucosa or bone meal, it is possible to solve problems such as stability, inclusion rate, and very high cost caused by using only existing bioactive substances directly. It is used as an excellent biodegradable carrier that can be applied to other organs such as ducts, blood vessels, liver, heart, urethra, and the like. Therefore, the present invention complexes the mammalian small intestinal submucosa or bone powder with a biodegradable polymer and uses it as a substitute for tissue regeneration.

Synthetic biodegradable polymer used in the present invention should be harmless to the human body and have properties that can be biodegraded within a desired period. Biodegradable synthetic polymers having such characteristics include chitin, chitosan, alginic acid, polylactic acid (PLA), polyglycolic acid (PGA), and lactic acid-glycolic acid copolymer (PLGA). At least one selected from polyhydroxybutyric acid, polycaprolactone, polyanhydride, and polyalkylcyano acrylate. These synthetic biodegradable polymers have been found to be naturally biodegradable after a certain period of time in the human body by hydrolysis and various enzymes in the human body [R. Langer and J. P. Vacanti, Science, 260, 920 (1993)].

Generally, salt extraction, gas expansion, solvent evaporation, phase separation, emulsion freeze drying, etc. may be used as a method for preparing a porous and biodegradable carrier. It was prepared by applying the salt extraction method in order to maximize.

Hereinafter, the manufacturing process of the artificial organ according to the present invention will be described in detail.

Since it is important to uniformly mix with the synthetic polymer solution, bone powder or small intestinal mucosa was dried and then micropowdered with a freeze mill. The finely divided bone powder or small intestinal submucosa powder was mixed with the polymer in the polymer solution (1.5 to 30 w / v% as methylene chloride (solvent)) in a weight ratio of 1: 0.1 to 1: 1.6, and then again to salt and 1: 4 to The carrier was prepared by mixing in a weight ratio of 1:16. At this time, since the pore size and porosity of the carrier are determined by the size and amount of the salt, the salt size is preferably 180 to 600 μm in diameter. In addition, when the mixing ratio of the bone powder or small intestinal submucosa powder and the polymer solution is less than 1: 0.1 there is a problem that can not take advantage of the bone powder and small intestinal submucosa, there is a problem of deteriorating the physical properties of the carrier when the ratio exceeds 1: 1.6, When the mixing ratio of the mixed solution and the salt is less than 1: 4, the porosity is less than 40%, and it is difficult to penetrate the inside of the cell. Meanwhile, the pore size, pore size distribution, porosity, and the like may be variously changed according to the type of artificial organ to be prepared, which is possible by controlling the content of the biodegradable polymer organic solvent and salt used.

The carrier according to the present invention is composed of a sponge-type biodegradable polymer having a pore size distribution and is implanted in the human body so that culture medium and cells can easily penetrate into the biodegradable carrier to allow for growth and recovery of tissue cells. The organs are slowly biodegraded by hydrolysis after a certain period of time, and there is no fear of being absorbed and accumulated in the human body, and there is a fear that the bioactive substances contained in the artificial organs may denature or water and other solvents may flow out during the manufacturing process. Bioactive substances in the small intestinal submucosa are sustained release and have excellent excellence in maintaining constant cell growth rate.

In addition, the artificial organs prepared using the above carriers are capable of growing and restoring tissue cells such as artificial blood vessels, artificial urethra, artificial small intestine, artificial bronchus, human space, artificial kidney, artificial cartilage, artificial bones, and artificial nerves. If it is a long term, it will apply.

Hereinafter, the present invention will be described in detail with reference to the following Examples, the present invention is not limited by the Examples.

Example 1

The small intestinal submucosa of pigs was separated, washed with saline, and lyophilized for 2 days at -55 ° C at 30 mTorr to remove moisture. To prepare a porous tissue engineering carrier, the dried small intestinal mucosa was micropowdered by grinding in a frozen state using a freeze mill [6700-115, Metuchem N.J., USA]. Micropowdered small intestinal submucosa and biodegradable polymer PLGA [75:25 mole ratio, Mw; 110,000, Beringer Ingelheim, Germany] After mixing the polymer in the solution in a weight ratio of 1: 1.0 and mixing the salt in a weight ratio of 1: 10 to the mixed solution, and put into a silicone mold of a disk form (diameter 10 mm, thickness 3 mm). The solvent was evaporated. After the solvent was completely removed, it was dissolved in distilled water to remove sodium chloride for 48 hours, and it was confirmed that there was no residual salt by using ion chromatography. The prepared porous carrier was lyophilized and stored in a vacuum oven.

The prepared porous carrier was photographed with a scanning electron microscope to observe the morphology of the pores [FIG. 1]. The pore size was uniformly formed, and the pore size and porosity could be controlled by the size and amount of salt. In addition, it was measured using a mercury porosimetry [Model AutoPore II 9220, Micromeritics Co., USA] to measure the distribution of pore size, non-porous area, average pore diameter, porosity. In particular, when the weight ratio of synthetic polymer and salt is 1: 8 or more, a carrier having a porosity of 90% or more could be prepared.

Example 2

In order to evaluate the wettability of the carrier in which the conventional synthetic polymer carrier and the small intestinal submucosa are complexed, the degree of wetting was observed by dropping the blue dye solution. This is not easy to regenerate the tissue because the cell does not penetrate the inside of the cell when the carrier is not wet in the culture medium when sowing. In order to compare with the PLGA carrier containing the small intestinal submucosa, a carrier made of PLGA itself was prepared and compared. At this time, the salt extraction method was used, the weight ratio of the total matrix and salt is 1:10 and the porosity was more than 90%. As shown in FIG. 3, the synthetic polymer carrier can be observed that the dye solution is not permeated and condensed on the surface. However, in the case of a carrier in which the small intestinal submucosa is complexed, the synthetic polymer carrier has a good surface property as a carrier for tissue engineering. Confirmed. Figure 4 shows the water absorption of the carrier. 20 ml of distilled water was added to the beaker, and the small intestinal submucosa was immersed by content, showing the water absorption rate (%) as the weight of the initial carrier and the weight ratio after impregnation. As a result, the water absorption rate of 600% or more was increased in the case of the carrier in which the small intestinal submucosa was complexed. As the content of the small intestinal submucosa increased, the water absorption rate also tended to increase.

Example 3

In order to observe the activity of various physiologically active substances, ie, growth factors or cytokines, included in the small intestinal submucosa used in the present invention, first, the small intestinal submucosa was finely ground with a homogenizer, and the extract was added to the culture solution. NIH / 3T3 fibroblasts were cultured using this culture and the proliferation rate of the cells was observed. As shown in Figure 5, it can be seen that the growth of the fibroblasts (b) cultured in the culture medium is added to the small intestinal submucosa extract compared to the fibroblasts (a) cultured in the existing culture medium. This is a fragmentary example that can be seen that there is a bioactive material that helps the proliferation of cells in the small intestinal submucosa.

Example 4

MTT assay was performed to determine the cell adhesion of PLGA synthetic polymer carrier according to the amount of intestinal submucosa. PLGA synthetic polymers and small intestinal submucosa were seeded in a carrier containing 1: 0.4, 1: 0.8, 1: 1.2 and 1: 1.6 cells at 5 × 10 ⁵ / mL density for 1 day, followed by 10 μL MTT solution. After incubating for 4 hours, the culture solution was removed and left overnight in 100 μL lysing buffer solution. The solution was collected and measured at 590 nm with a spectrophotometer. Figure 6 shows the measured value graphically. As the intestinal submucosa increases in content, the cell adhesion is better. This may be due to the extracellular matrix (e.g. collagen, proteoglycan, sticky protein) that make up the small intestinal submucosa.

Example 5

For the preparation of histological organs of a carrier complexed with small intestinal mucosa and PLGA synthetic polymer, various cells were seeded and adhered for 2 hours, and then implanted subcutaneously in immunodeficient nude mice. As a result of observing this after 4 weeks, as shown in FIG. 7, it was visually confirmed that the size of the implanted carrier was significantly reduced in the case of PLGA carrier (uncombined small intestinal submucosa). This indicates that the cells adhere smoothly and proliferate in the carrier to produce extracellular matrix. To confirm this, the two carriers that were transplanted were examined by H & E (hematoxylin and eosin) staining. 8 shows the result of staining the PLGA polymer carrier (a) and the result of the carrier of the submucosal tissue complex (b). In the case of the PLGA polymer carrier, the cells were not deeply inserted and the density of the cells was significantly low. Carrier complexed with submucosal tissue had a high density of cells and contained cells deep inside. This indicates that the carrier presented in the present invention is excellent in tissue engineering.

Example 6

Production of various organs using a carrier in which the small intestinal submucosal tissue is complexed can be presented. In this embodiment, the method of preparing artificial tissue cartilage by seeding chondrocytes is as follows.

Human chondrocytes were isolated and cultured in a cell incubator. The number of cultured cells was 5 × 10 ⁵ / mL concentration so that the small intestinal submucosa was seeded on the complex carrier. After 2 hours of incubation in the cell culture, they were implanted subcutaneously in immunodeficient nude mice. FIG. 9 shows safranin-O staining to confirm glycosaaminoglycans of cartilage tissue, and it can be seen that cartilage tissue is generated from the carrier of the submucosal tissue and PLGA polymer complex of the present invention.

Example 7

In this embodiment, the method for producing artificial tissue bone by sowing bone cells is as follows.

Human myeloid mesenchymal cells were isolated and cultured in a cell culture phase. Cultured cells were differentiated into bone cells and confirmed this. The number of cultured cells was 5 × 10 ⁵ / mL concentration so that the small intestinal submucosa was seeded on the complex carrier. This was incubated for 2 hours in a cell culture period, and then transplanted subcutaneously in immunodeficient nude mice. 10 is a Bon-Kussa staining for identifying the bone tissue, it can be observed that the cells seeded in the small intestinal submucosa and PLGA polymer carriers presented in the present invention to form a new bone calcium.

Example 8

The present embodiment relates to the preparation of bone flour, a method for preparing a carrier in which bone flour and synthetic polymer are combined, and tissue engineering bone regeneration using the same. Specific methods for this are as follows.

Bone tissue isolated from mammals was treated with strong acid to remove inorganic components and powdered. Bone powder from which minerals were removed was mixed with PLGA solution, which is a biodegradable polymer, and sodium chloride in a weight ratio of 1: 0.4, 0.8, 1.2, and 1.6, and then placed in a mold having a disk shape (diameter 10 mm, thickness 3 mm) to evaporate the solvent. The size of the pore was adjusted by the size of sodium chloride, and the porosity was adjusted by the amount thereof. After the solvent was completely removed, it was dissolved in distilled water to remove sodium chloride and dissolved for 48 hours, and it was confirmed that there was no residual salt by using ion chromatography. The prepared porous carrier was lyophilized and stored in a vacuum oven.

The prepared porous carrier was photographed with a scanning electron microscope to observe the shape of the pores [FIG. 2]. The pore size was uniformly formed, and the pore size and porosity could be controlled by the size and amount of salt. In addition, a mercury porosimetry (Model AutoPore II 9220, Micromeritics Co., USA) was used to measure its pore size distribution, specific pore area, average pore diameter, and porosity.

Example 9

Production of various organs can be suggested using PLGA carriers in which bone meal is complexed. In this embodiment, a method of producing artificial tissue bone by sowing bone cells is as follows.

Human bone marrow mesenchymal cells were isolated on PLGA carriers in which the bone marrow of mammals was mixed and seeded with cells differentiated into bone cells at a concentration of 5 × 10 ⁵ / mL. This was incubated for 2 hours in a cell culture period, and then transplanted subcutaneously in immunodeficient nude mice. 11 confirmed that the cells seeded in the PLGA carrier complexed with bone powder secrete the substrate and form new bone tissue.

As described above, the natural / synthetic polymer hybrid carrier according to the present invention has a suitable pore size, so that material transfer such as nutrient solution, oxygen, carbon dioxide, etc. is effectively carried out between blood and skin, and the connection of this multi-space is biodegradable. Because it is made of a polymer, it is easy to attach and grow vascular endothelial cells or skin tissue cells in the human body, and after the growth of these cells is completed, it is naturally biodegradable and absorbed by the body, which is superior to conventional artificial organ materials. Have

In addition, the carrier is excellent in water absorption, flexibility is increased, the processability is very good, and the growth, differentiation and growth of the cell by the various growth factors and cytokines contained in the tissue itself has the advantage that the conventional Biodegradable polymer carriers, which are indispensable in the development of tissue engineering artificial organs, can be functionalized and have much easier and superior economic advantages than conventional methods.

Claims (7)

A carrier for tissue engineering, characterized in that the biomaterial of the small intestine submucosa or bone powder of a mammal is prepared by complexing with a synthetic polymer. According to claim 1, wherein the mammal is a tissue engineering carrier, characterized in that selected from cows, pigs, rabbits, rats. The method of claim 1, wherein the synthetic polymer is chitin, chitosan, alginic acid, polylactic acid (PLA), polyglycolic acid (PGA), lactic acid-glycolic acid copolymer (PLGA), poly A carrier for tissue engineering, characterized in that at least one selected from hydroxybutyric acid, polycaprolactone, polyanhydride and polyalkylcyano acrylate. 1) lyophilizing and pulverizing the small intestinal submucosa or bone powder of a mammal; 2) mixing the micropowdered small intestinal submucosa or bone powder with a synthetic polymer solution in a weight ratio of 1: 0.4 to 1: 1.6; 3) mixing a salt in a weight ratio of 1: 4 to 1:16 to the mixed solution, and then evaporating the solvent; And 4) dissolving the residue of 3) in distilled water to completely remove the salt; Method for producing a tissue engineering carrier, characterized in that it comprises a. The method of claim 4, wherein the mammal is selected from cows, pigs, rabbits, and rats. The method of claim 4, wherein the synthetic polymer is chitin, chitosan, alginic acid, polylactic acid (PLA), polyglycolic acid (PGA), lactic acid-glycolic acid copolymer (PLGA), poly Method for producing a tissue engineering carrier, characterized in that at least one selected from hydroxybutyric acid (polyhydroxybutyric acid), polycaprolactone, polyanhydride and polyalkylcyano acrylate (polyalkylcyano acrylate). A porous, biodegradable artificial organ, which is prepared using the carrier of claim 1.