KR20210078793A - Bio ink composition, preparing method for 3-dimension scafford of skin and soft tissue regeneration and 3-dimension scafford thereby - Google Patents

Abstract

The present invention relates to a bio-ink composition, a method for producing a three-dimensional scaffold that can be used for skin and soft tissue regeneration, and a three-dimensional scaffold produced thereby. The three-dimensional scaffold according to an embodiment of the present invention comprises: a biodegradable material; growth factor substances; and antibiotics.

Description

Bio-ink composition, method for producing a three-dimensional scaffold that can be used for skin and soft tissue regeneration, and a three-dimensional scaffold prepared thereby SCAFFORD THEREBY}

The present invention relates to a bio-ink composition, a method for manufacturing a three-dimensional scaffold that can be used for skin and soft tissue regeneration, and a three-dimensional scaffold prepared thereby.

A cell carrier (scaffold) is a biomaterial substitute that structurally supports a part of a damaged body, and is an artificial tissue and organ that can directly replace an organ until the tissue or organ is regenerated. Or it serves as a prosthesis.

The structure of the cell carrier for this role must have mechanical strength and characteristics that can coexist with the tissues in the body and perform its role, and must be manufactured in a form with the biological properties of the human body that can restore and promote tissue. . It is manufactured using biodegradable polymers as the main material for cell carriers. Recently, in order to improve biological functions such as cell culture, proliferation, and differentiation, protein-based (collagen, alginate, chitosan, silk-fibroin) similar to the properties of actual tissues (collagen, alginate, chitosan, silk-fibroin) , and gelatin etc.) are increasing cases of using hydrogels.

Protein-based hydrogels are widely used as bio-inks for 3D bioprinting due to the characteristics of proteins (biodegradability, biocompatibility, cytophilicity), and further research is actively using them as 3D protein scaffolds necessary for the regeneration of body organs such as bone and cartilage. is in progress However, gelatin, which has excellent biocompatibility, has a weakness to be used as a structural and physical support for bone due to its physical property of melting at body temperature (37 °C). To compensate for this, a three-dimensional shape is produced by crosslinking with genipin or glutaraldehyde, which is a chemical crosslinker. However, since the crosslinker is toxic, biocompatibility is very unstable.

Recently, by crosslinking not only collagen but also various natural polymers in various ways to improve mechanical strength, which has been pointed out as a disadvantage in the past, has been used not only for bone regeneration but also for the regeneration of various soft tissues. However, for application to tissue engineering, it has many problems such as low viscosity, mechanical properties, degradation rate, structure, and morphology, etc. Do.

SUMMARY OF THE INVENTION The present invention is to solve the above problems, and an object of the present invention is a bio-ink composition having biological activity including immunology for soft tissue clinical applications, skin and soft tissue An object of the present invention is to provide a method for producing a three-dimensional scaffold that can be used for regeneration, and a three-dimensional scaffold prepared thereby.

However, the problems to be solved by the present invention are not limited to those mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

A bio-ink composition according to an embodiment of the present invention includes a biodegradable material; and growth factors and antibiotics.

According to one embodiment, the biodegradable material is, collagen (collagen), alginic acid (alginic acid), albumin (albumin), gelatin (gelatin), chitosan (chitosan), silk fibroin (silk fibroin), polypeptide (polypeptide) , heparin, alginate, dextran, chondroitin sulfate, dextran sulfate, acacia gum, tragacanthin, pectin , guar gum, hyaluronic acid, sodium carboxymethyl dextran, carboxymethyl cellulose, peptide, oligopeptide, agar, carrageenan (carrageenan), Galactomannans, Xanthan, Beta-Cyclodextrin, Amylose (Soluble Starch), Fibrin, Pullulan and Tertiary It may include at least one selected from the group consisting of amine starch ether).

According to an embodiment, the biodegradable material may be 1 wt% to 50 wt% of the bio-ink composition.

According to an embodiment, the growth factor material is adrenomedullin, angiopoietin, autocrine motility factor, bone morphogenetic proteins, ciliary neurotrophic Ciliary neurotrophic factor, Leukemia inhibitory factor, Interleukin-1, Interleukin-2, Interleukin-3, Interleukin-4 ), Interleukin-5, Interleukin-6, Interleukin-7, Colony-stimulating factors, Macrophage colony-stimulating factor ), Granulocyte colony-stimulating factor, Granulocyte macrophage colony-stimulating factor, Epidermal growth factor, Ephrin A1, Ephrin A2 ( Ephrin A2), Ephrin A3 (Ephrin A3), Ephrin A4 (Ephrin A4), Ephrin A5 (Ephrin A5), Ephrin B1 (Ephrin B1), Ephrin B2 (Ephrin B2), Ephrin B3 (Ephrin B3) ), erythropoietin, fibroblast growth factor 1-23, bovine somatotrophin, glial cell line-derived neurotrophic factor, Neurturin, Persephin, Artemin (A) rtemin), Growth differentiation factor-9, Hepatocyte growth factor, Hepatoma-derived growth factor Insulin, Insulin-like growth factors , Insulin-like growth factor-1 (Insulin-like growth factor-1), insulin-like growth factor-2 (Insulin-like growth factor-2), interleukins (Interleukins), keratinocyte growth factor (Keratinocyte growth factor), cell migration Migration-stimulating factor, macrophage-stimulating protein, myostatin, neuregulin 1-4, neurotrophins, brain-derived nerve management Brain-derived neurotrophic factor, Nerve growth factor, Neurotrophin-3, Neurotrophin-4, Placental growth factor, Renalase (Renalase), T-cell growth factor (T-cell growth factor), thrombopoietin, transforming growth factor (Transforming growth factor), transforming growth factor alpha (Transforming growth factor alpha), transforming growth factor At least one selected from the group consisting of beta (Transforming growth factor beta), tumor necrosis factor-alpha, vascular endothelial growth factor, and wnt signaling pathway (Wnt Signaling Pathway) to include can be

According to one embodiment, the growth factor material may be 0.0001 wt% to 30 wt% of the bio-ink composition.

According to one embodiment, the antibiotic component, penicillin (Penicillin), nafcillin (Nafcillin), ampicillin / sulbactam (Ampicillin / sulbactam), amoxicillin (Amoxicillin), cefazolin (Cefazolin), cephalexin (Cephalexin), cepra Cephradine, Cefadroxil, Clindamycin, Doxycycline, Trimethoprim/sulfamethoxazole, Vancomycin, Linezolid, Bacitracin (bacitracin), gentamicin (Gentamicin), fusidic acid (Fusidic acid), may be one containing at least one of mupirocin (mupirocin).

According to an embodiment, the antibiotic component may be 0.00000001 wt% to 30 wt% of the bio-ink composition.

According to an embodiment, the growth factor may be 0.0001% to 30% by weight of the bio-ink composition.

According to an embodiment, the bio-ink composition may further include a crosslinking agent.

According to one embodiment, the crosslinking agent, tannic acid (tannic acid), genipin (genipin), transglutaminase ((Moo Glo TI) Transglutaminase), ethyldimethylaminopropylcarbodiimide [1-ethyl-3 (3-dimethyl aminopropyl) carbodiimide; EDC], hydroxysuccinimide (N-hydroxysuccinimide; NHS), dimethylaminopropyl ethylcarbodiimide hydrochloride (EDAC), hydroxybenzotriazole (hydroxybenzotriazole) and glutaraldehyde (Glutaraldehyde) consisting of It may include at least one selected from the group.

According to an embodiment, the crosslinking agent may be 0.00001 wt% to 20 wt% of the bio-ink composition.

A method of manufacturing a three-dimensional scaffold according to another embodiment of the present invention includes the steps of preparing a bio-ink composition according to an embodiment of the present invention; outputting the bio-ink composition as a three-dimensional structure using 3D printing; and adding a crosslinking agent to the three-dimensional structure and curing it.

According to an embodiment, preparing the bio-ink composition includes: preparing a biodegradable material; dialysis of the biodegradable material; centrifuging the dialyzed biodegradable material; and mixing a growth factor material and an antibiotic component with the centrifuged biodegradable material;

According to one embodiment, the step of outputting the three-dimensional structure using 3D printing the bio-ink composition comprises: outputting the bio-ink composition in a cooling jacket of 0 °C to 20 °C; and cooling on a 0 °C to -100 °C cooling stage.

According to one embodiment, the crosslinking agent, tannic acid (tannic acid), genipin (genipin), transglutaminase ((Moo Glo TI) Transglutaminase), ethyldimethylaminopropylcarbodiimide [1-ethyl-3 (3-dimethyl aminopropyl) carbodiimide; EDC], hydroxysuccinimide (N-hydroxysuccinimide; NHS), dimethylaminopropyl ethylcarbodiimide hydrochloride (EDAC), hydroxybenzotriazole (hydroxybenzotriazole) and glutaraldehyde (Glutaraldehyde) consisting of It may include at least one selected from the group.

A three-dimensional scaffold according to another embodiment of the present invention is manufactured by the method for manufacturing a three-dimensional scaffold according to an embodiment of the present invention.

According to an embodiment, the three-dimensional scaffold has a compressive modulus of 0.00001 MPa to 10 MPa, a surface roughness (Ra) of 10 nm to 10 μm, and water-absorbability. It may be 0.1% or more.

The bio-ink composition according to an embodiment of the present invention can mimic the mechanical properties of a target soft tissue according to cross-linking conditions, and contains growth factors and antibiotics appropriately to protect from the risk of secondary infection when transplanted into the body It is possible to manufacture a three-dimensional scaffold with improved tissue repair and blood vessel formation and improved immunity due to improved tissue repair and blood vessel formation.

A 3D printing-based biodegradable scaffold manufacturing process can be established by the 3D scaffold manufacturing method according to an embodiment of the present invention, and through this, a porous biodegradable skin and soft tissue application implant material can be developed. In particular, it is possible to minimize the change in viscosity according to the temperature of the hydrogel monomer through two-step temperature control, and it can be produced and supplied with high reproducibility for long-term regeneration.

When the three-dimensional scaffold according to an embodiment of the present invention is used as a scaffold for bone regeneration, not only mechanical strength close to an actual organ but also human-friendly biological properties can be expected compared to the conventional scaffold.

1 is a flowchart showing the sequence of a method for manufacturing a three-dimensional scaffold according to an embodiment of the present invention.
FIG. 2 is a flowchart illustrating detailed steps of preparing the bio-ink composition of FIG. 1 .
3 is a flowchart illustrating detailed steps of the output step of the three-dimensional structure of FIG. 1 .
4 is a schematic diagram illustrating a printer in which a temperature control device capable of controlling the temperature of a printer head and a stage and a sensor are combined during the manufacturing process of the three-dimensional scaffold according to an embodiment of the present invention.
5 is an image of a sample and a step of outputting a three-dimensional structure by controlling the temperature of two steps during the manufacturing process of the three-dimensional scaffold according to an embodiment of the present invention.
6 is an image of a sample in which antibiotic components and growth factors are released from a three-dimensional structure fabricated during the manufacturing process of a three-dimensional scaffold according to an embodiment of the present invention.

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. In describing the present invention, if it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the gist of the present invention, the detailed description thereof will be omitted. In addition, the terms used in this specification are terms used to properly express a preferred embodiment of the present invention, which may vary depending on the intention of a user or operator or a custom in the field to which the present invention belongs. Accordingly, definitions of these terms should be made based on the content throughout this specification. Like reference numerals in each figure indicate like elements.

Throughout the specification, when a member is said to be located "on" another member, this includes not only a case in which a member is in contact with another member but also a case in which another member exists between the two members.

Throughout the specification, when a part "includes" a certain component, it means that other components may be further included, rather than excluding other components.

Hereinafter, the bio-ink composition of the present invention, a method for producing a three-dimensional scaffold that can be used for skin and soft tissue regeneration, and a three-dimensional scaffold prepared thereby will be described in detail with reference to Examples and drawings. However, the present invention is not limited to these examples and drawings.

As used herein, the term "scaffold" may refer to a material that replaces a part of an organ or tissue damaged in vivo and can supplement or replace its function. In particular, the three-dimensional scaffold may include a biodegradable material, which may be completely decomposed and disappear in vivo after the scaffold is maintained until it sufficiently performs its function and role.

A bio-ink composition according to an embodiment of the present invention includes a biodegradable material; and growth factors and antibiotics.

According to an embodiment, the biodegradable material is made of a homopolymer or a copolymer, and forms a structurally stable three-dimensional network structure with little fluidity due to external history, such as a covalent bond, hydrogen bond, van der It is formed by several factors, such as a Bals bond or physical aggregation.

According to one embodiment, the biodegradable material is, collagen (collagen), alginic acid (alginic acid), albumin (albumin), gelatin (gelatin), chitosan (chitosan), silk fibroin (silk fibroin), polypeptide (polypeptide) , heparin, alginate, dextran, chondroitin sulfate, dextran sulfate, acacia gum, tragacanthin, pectin , guar gum, hyaluronic acid, sodium carboxymethyl dextran, carboxymethyl cellulose, peptide, oligopeptide, agar, carrageenan (carrageenan), Galactomannans, Xanthan, Beta-Cyclodextrin, Amylose (Soluble Starch), Fibrin, Pullulan and Tertiary It may include at least one selected from the group consisting of amine starch ether).

According to an embodiment, the biodegradable material may be 1 wt% to 50 wt% of the bio-ink composition. If the biodegradable material is less than 1% by weight of the bio-ink composition, a problem of fabricating the structure may occur during printing due to low viscosity, and if it is more than 50% by weight, the viscosity may be high during printing and thus a problem of ejection may occur.

According to an embodiment, the growth factor material is adrenomedullin, angiopoietin, autocrine motility factor, bone morphogenetic proteins, ciliary neurotrophic Ciliary neurotrophic factor, Leukemia inhibitory factor, Interleukin-1, Interleukin-2, Interleukin-3, Interleukin-4 ), Interleukin-5, Interleukin-6, Interleukin-7, Colony-stimulating factors, Macrophage colony-stimulating factor ), Granulocyte colony-stimulating factor, Granulocyte macrophage colony-stimulating factor, Epidermal growth factor, Ephrin A1, Ephrin A2 ( Ephrin A2), Ephrin A3 (Ephrin A3), Ephrin A4 (Ephrin A4), Ephrin A5 (Ephrin A5), Ephrin B1 (Ephrin B1), Ephrin B2 (Ephrin B2), Ephrin B3 (Ephrin B3) ), erythropoietin, fibroblast growth factor 1-23, bovine somatotrophin, glial cell line-derived neurotrophic factor, Neurturin, Persephin, Artemin (A) rtemin), Growth differentiation factor-9, Hepatocyte growth factor, Hepatoma-derived growth factor Insulin, Insulin-like growth factors , Insulin-like growth factor-1, insulin-like growth factor-2 (Insulin-like growth factor-2), interleukins, keratinocyte growth factor (Keratinocyte growth factor), cell migration Migration-stimulating factor, Macrophage-stimulating protein, Myostatin, Neuregulin 1-4, Neurotrophins, brain-derived nerve management Brain-derived neurotrophic factor, Nerve growth factor, Neurotrophin-3, Neurotrophin-4, Placental growth factor, Renalase (Renalase), T-cell growth factor (T-cell growth factor), thrombopoietin, transforming growth factor (Transforming growth factor), transforming growth factor alpha (Transforming growth factor alpha), transforming growth factor At least one selected from the group consisting of beta (Transforming growth factor beta), tumor necrosis factor-alpha, vascular endothelial growth factor, and wnt signaling pathway (Wnt Signaling Pathway) to include can be

According to one embodiment, the growth factor material may be 0.0001 wt% to 30 wt% of the bio-ink composition. If the growth factor material is less than 0.0001 wt% of the bio-ink composition, the effect of the growth factor on the target tissue is insignificant and may be inactivated, and if the growth factor material is more than 30 wt%, abnormal cell growth or genetic modification occurs, resulting in chemical and As a biochemical stimulatory factor, the activity and differentiation of cells and tissues may be abnormal.

According to one embodiment, the antibiotic component, penicillin (Penicillin), nafcillin (Nafcillin), ampicillin / sulbactam (Ampicillin / sulbactam), amoxicillin (Amoxicillin), cefazolin (Cefazolin), cephalexin (Cephalexin), cepra Cephradine, Cefadroxil, Clindamycin, Doxycycline, Trimethoprim/sulfamethoxazole, Vancomycin, Linezolid, Bacitracin (bacitracin), gentamicin (Gentamicin), fusidic acid (Fusidic acid), may be one containing at least one of mupirocin (mupirocin).

According to an embodiment, the antibiotic component may be 0.00000001 wt% to 30 wt% of the bio-ink composition. If the antibiotic content is less than 0.0001 wt% of the bio-ink composition, it cannot perform an antibiotic role and thus cannot prevent bacterial and bacterial infection, and if it exceeds 30 wt%, excessive antibiotic ingredients not only interfere with the growth of cells, but also target tissues or tissues. Adverse effects may occur in surrounding tissues and organs.

According to an embodiment, the growth factor may be 0.0001% to 30% by weight of the bio-ink composition.

According to an embodiment, the bio-ink composition may further include a crosslinking agent.

According to one embodiment, the crosslinking agent, tannic acid (tannic acid), genipin (genipin), transglutaminase ((Moo Glo TI) Transglutaminase), ethyldimethylaminopropylcarbodiimide [1-ethyl-3 (3-dimethyl aminopropyl) carbodiimide; EDC], hydroxysuccinimide (N-hydroxysuccinimide; NHS), dimethylaminopropyl ethylcarbodiimide hydrochloride (EDAC), hydroxybenzotriazole (hydroxybenzotriazole) and glutaraldehyde (Glutaraldehyde) consisting of It may include at least one selected from the group.

According to an embodiment, the crosslinking agent may be 0.00001 wt% to 20 wt% of the bio-ink composition. If the cross-linking agent is less than 0.00001 wt% of the bio-ink composition, there may be a problem in that mechanical properties cannot be improved when manufacturing a three-dimensional scaffold, and if it is more than 20 wt%, it is difficult to use as a soft tissue graft material due to excessive mechanical properties there may be

The bio-ink composition according to an embodiment of the present invention can mimic the mechanical properties of a target soft tissue according to cross-linking conditions, and contains growth factors and antibiotics appropriately to protect from the risk of secondary infection when transplanted into the body It is possible to manufacture a three-dimensional scaffold with improved tissue repair and blood vessel formation.

A method of manufacturing a three-dimensional scaffold according to another embodiment of the present invention includes the steps of preparing a bio-ink composition according to an embodiment of the present invention; outputting the bio-ink composition as a three-dimensional structure using 3D printing; and adding a crosslinking agent to the three-dimensional structure and curing it.

1 is a flowchart showing a sequence of a method of manufacturing a three-dimensional scaffold according to an embodiment of the present invention.

Referring to FIG. 1 , the method for manufacturing a three-dimensional scaffold according to an embodiment of the present invention includes a bio-ink composition preparation step 110 , a three-dimensional structure output step 120 , and a three-dimensional structure curing step 130 . include

According to one embodiment, the bio-ink composition preparation step 110, a biodegradable material according to an embodiment of the present invention; growth factor substances; and an antibiotic component; to prepare a bio-ink composition comprising.

According to an embodiment, preparing the bio-ink composition includes: preparing a biodegradable material; dialysis of the biodegradable material; centrifuging the dialyzed biodegradable material; and mixing a growth factor material and an antibiotic component with the centrifuged biodegradable material;

FIG. 2 is a flowchart illustrating detailed steps of preparing the bio-ink composition of FIG. 1 .

Referring to FIG. 2 , the bio-ink composition preparation step according to an embodiment of the present invention includes a biodegradable material preparation step 112 , a biodegradable material dialysis step 114 , a biodegradable material centrifugation step 116 and growth and mixing the factor material and the antibiotic component (118).

According to one embodiment, the biodegradable material preparation step 112 is a step of preparing the biodegradable material described above. Since specific types of biodegradable materials have been described above, overlapping descriptions will be omitted.

According to one embodiment, the biodegradable material is put in DPBS (Dulleco's phosphate buffer) and dissolved at a temperature of 40 ° C. to 60 ° C. for 30 minutes to 3 hours, and then, the biodegradable material - methacrylic anhydride (Methacrylic) in DPBS solution anhydride) may be mixed and the reaction proceeded for 2 to 4 hours.

According to one embodiment, in the dialysis step 114 of the biodegradable material, the opaque solution in which the reaction has progressed is put in 3 to 5 times the DBPS solution to which it was first put to terminate the reaction, and then divided into dialysis tubing. After that, it may be to be dialyzed for one week using DI water at 30 °C to 50 °C.

According to one embodiment, the centrifugation step 116 of the biodegradable material may be centrifuging the dialyzed solution. After centrifugation, it may be freeze-dried.

According to one embodiment, in the step 118 of mixing the growth factor material and the antibiotic component, the growth factor material and the antibiotic component may be mixed with the centrifuged biodegradable material. Since the specific types of growth factor materials and antibiotics have been described above, overlapping descriptions will be omitted.

According to one embodiment, the step of outputting the three-dimensional structure using 3D printing the bio-ink composition comprises: outputting the bio-ink composition in a cooling jacket of 0 °C to 20 °C; and cooling on a 0 °C to -100 °C cooling stage.

According to one embodiment, the outputting step 120 of the three-dimensional structure is to output the bio-ink composition as a three-dimensional structure using 3D printing.

3 is a flowchart illustrating detailed steps of the output step of the three-dimensional structure of FIG. 1 .

Referring to FIG. 3 , the 3D structure output step 120 may include a bio-ink composition output step 122 and a cooling step 124 .

4 is a schematic diagram illustrating a printer in which a temperature control device capable of controlling the temperatures of a printer head and a stage and a sensor are combined during the manufacturing process of a three-dimensional scaffold according to an embodiment of the present invention.

Referring to FIG. 4 , in manufacturing the three-dimensional scaffold according to an embodiment of the present invention, the step of outputting the three-dimensional structure includes a cooling jacket made of a peltier material and a cooling stage ( It may be to use a 3D printer combined with a cooling stage).

5 is an image of a sample and a step of outputting a three-dimensional structure by controlling the temperature of two steps during the manufacturing process of the three-dimensional scaffold according to an embodiment of the present invention.

Referring to FIG. 5 , in manufacturing the three-dimensional scaffold according to an embodiment of the present invention, the manufactured three-dimensional structure provides an antibiotic component to protect from secondary infection after transplantation, and angiogenesis and tissue repair. It may be to use a possible growth factor.

6 is an image of a sample in which antibiotics and growth factors are released from a three-dimensional structure fabricated during the manufacturing process of a three-dimensional scaffold according to an embodiment of the present invention.

Referring to FIG. 6 , in manufacturing the three-dimensional scaffold according to an embodiment of the present invention, the manufactured three-dimensional structure provides an antibiotic component that protects from secondary infection after transplantation, and aids in angiogenesis and tissue repair. It may be to use a growth factor that can be given.

According to one embodiment, it may be to apply a two-stage cooling system in order to minimize the change in viscosity according to the temperature of the biodegradable material.

According to an embodiment, the bio-ink composition output step 122 is to output the bio-ink composition in a cooling jacket at 0° C. to 20° C. using 3D printing of the bio-ink composition.

According to one embodiment, the cooling step 124 is cooling on a 0 °C to -100 °C cooling stage.

According to one embodiment, the curing step 130 of the crosslinking agent is to add a crosslinking agent to the three-dimensional structure and harden.

According to one embodiment, the crosslinking agent, tannic acid (tannic acid), genipin (genipin), transglutaminase ((Moo Glo TI) Transglutaminase), ethyldimethylaminopropylcarbodiimide [1-ethyl-3 (3-dimethyl aminopropyl) carbodiimide; EDC], hydroxysuccinimide (N-hydroxysuccinimide; NHS), dimethylaminopropyl ethylcarbodiimide hydrochloride (EDAC), hydroxybenzotriazole (hydroxybenzotriazole) and glutaraldehyde (Glutaraldehyde) consisting of It may include at least one selected from the group.

A 3D printing-based biodegradable scaffold manufacturing process can be established by the 3D scaffold manufacturing method according to an embodiment of the present invention, and through this, a porous biodegradable soft tissue reconstruction/new implant material can be developed. In particular, it is possible to minimize the change in viscosity according to the temperature of the hydrogel monomer through two-step temperature control, and it can be produced and supplied with high reproducibility for long-term regeneration.

A three-dimensional scaffold according to another embodiment of the present invention is manufactured by the method for manufacturing a three-dimensional scaffold according to an embodiment of the present invention.

According to an embodiment, the three-dimensional scaffold may have a diameter of 25 μm to 1 mm in one strand. If the diameter is less than 25 μm, there may be a difficult problem in implementing a shape with a 3D printer using a natural polymer material, and if it is more than 2 mm, there may be a difficult problem in implementing the shape with a 3D printer.

According to an embodiment, the three-dimensional scaffold may have a pore size of 1 μm to 2 mm. If the diameter is less than 1 μm, there may be a problem outside the lowest resolution that can implement a shape with a 3D printer, and if it is more than 2 mm, there may be a problem that the tissue repair speed may be slow.

According to an embodiment, the three-dimensional scaffold may have a porosity of 1% to 90%. If the porosity is less than 1%, there may be a problem that the movement and penetration of cells may be limited, and if it is more than 90%, there may be a problem in that the mechanical properties of the target tissue cannot be simulated.

According to an embodiment, the three-dimensional scaffold may have a compressive modulus of 0.00001 MPa to 10 MPa. When the compressive coefficient is less than 0.00001 MPa, there may be a problem in mechanical strength during implantation due to low mechanical strength, and if it exceeds 10 MPa, there may be a problem in inhibiting biodegradability due to high mechanical strength.

According to an embodiment, the three-dimensional scaffold may have a surface roughness (Ra) of 10 nm to 10 μm. If the surface roughness is less than 10 nm, there may be a limited problem in the initial adhesion of cells, and if it is more than 10 μm, there may be a problem in that the movement of cells may be limited.

According to an embodiment, the three-dimensional scaffold may have a water-absorbability of 0.1% or more. If the water absorption rate is less than 0.1%, there may be a problem that may limit the initial adhesion of cells.

According to one embodiment, a biomaterial may be immobilized on the three-dimensional scaffold. As such a biomaterial, at least one selected from the group consisting of growth factors, drugs, amino acids, proteins, lipids, carbohydrates, carbohydrates, nucleic acids, enzymes, minerals, hormones and antigens may be used. The three-dimensional scaffold may be usefully used for movement and delivery of biomaterials.

The three-dimensional scaffold according to an embodiment of the present invention is implanted in vivo for the purpose of tissue regeneration, and the scaffold made of a biodegradable polymer can be mainly used for bone tissue and cartilage tissue. At this time, in order to promote tissue regeneration, cells such as stem cells penetrate and adhere to the inside of the scaffold to proliferate and/or differentiate, and growth factors including nutrients for cell entry and exit as well as growth and/or differentiation thereof It is important that the supply of materials such as such is easy to enter and exit. Therefore, it is preferable that the scaffold used for this has a size and shape suitable for the site to be implanted, as well as an open pore of an appropriate size for passing from its surface to the inside. In the present invention, a scaffold is manufactured by printing in a 3D modeled form, but using a bio-ink composition based on biodegradable materials, antibiotics, and growth factors, etc., when printing through a high-temperature nozzle, it has the input appearance and mechanical properties It is characterized by having pores connected to the inside from the surface as well as improving it.

When the three-dimensional scaffold according to an embodiment of the present invention is used as a scaffold for bone regeneration, not only high mechanical strength but also human-friendly biological properties can be expected compared to the conventional scaffold.

As described above, although the embodiments have been described with reference to the limited embodiments and drawings, various modifications and variations are possible from the above description by those skilled in the art. For example, even if the described techniques are performed in an order different from the described method, and/or the described components are combined or combined in a different form from the described method, or replaced or substituted by other components or equivalents Appropriate results can be achieved. Therefore, other implementations, other embodiments, and equivalents to the claims are also within the scope of the following claims.

Claims (16)

biodegradable material;
growth factor substances; and
antibiotics;
comprising,
Bio-ink composition.
According to claim 1,
The biodegradable material, the biodegradable material, collagen (collagen), alginic acid (alginic acid), albumin (albumin), gelatin (gelatin), chitosan (chitosan), silk fibroin (silk fibroin), polypeptide (polypeptide) , heparin, alginate, dextran, chondroitin sulfate, dextran sulfate, acacia gum, tragacanthin, pectin , guar gum, hyaluronic acid, sodium carboxymethyl dextran, carboxymethyl cellulose, peptide, oligopeptide, agar, carrageenan (carrageenan), Galactomannans, Xanthan, Beta-Cyclodextrin, Amylose (Soluble Starch), Fibrin, Pullulan and Tertiary amine starch ether) comprising at least any one selected from the group consisting of,
Bio-ink composition.
According to claim 1,
The biodegradable material will be 1% to 50% by weight of the bio-ink composition,
Bio-ink composition.
According to claim 1,
The growth factor material is adrenomedullin, angiopoietin, autocrine motility factor, bone morphogenetic proteins, ciliary neurotrophic factor. , Leukemia inhibitory factor, Interleukin-1, Interleukin-2, Interleukin-3, Interleukin-4, Interleukin-5 ( Interleukin-5), Interleukin-6, Interleukin-7, Colony-stimulating factors, Macrophage colony-stimulating factor, granulocyte colony stimulating factor (Granulocyte colony-stimulating factor), Granulocyte macrophage colony-stimulating factor, Epidermal growth factor, Ephrin A1, Ephrin A2, Ephrin Ephrin A3, Ephrin A4, Ephrin A5, Ephrin B1, Ephrin B2, Ephrin B3, Erythropoietin , Fibroblast growth factor 1-23, bovine somatotrophin, glial cell line-derived neurotrophic factor, Nuturin, Persephin, Artemin, Growth differentiation Growth differentiation factor-9, Hepatocyte growth factor, Hepatoma-derived growth factor Insulin, Insulin-like growth factor, Insulin-like growth factor -1 (Insulin-like growth factor-1), insulin-like growth factor-2 (Insulin-like growth factor-2), interleukins, keratinocyte growth factor, cell migration stimulating factor (Migration- stimulating factor), granulocyte macrophage growth factor (Macrophage-stimulating protein), Myostatin, Neuregulin 1-4, Neurotrophins, Brain-derived neurotrophic factor), nerve growth factor (Nerve growth factor), neurotrophin 3 (Neurotrophin-3), neurotrophin 4 (Neurotrophin-4), placental growth factor (Placental growth factor), renalase (Renalase), T Cell growth factor (T-cell growth factor), thrombopoietin (Thrombopoietin), transforming growth factor (Transforming growth factor), transforming growth factor alpha (Transforming growth factor alpha), transforming growth factor beta (Transforming growth factor) beta), tumor necrosis factor alpha (Tumor necrosis factor-alpha), vascular endothelial growth factor (Vscular endothelial growth factor) and wnt signaling pathway (Wnt Signaling Pathway) comprising at least one selected from the group consisting of,
Bio-ink composition.
According to claim 1,
The growth factor material will be 0.0001 wt% to 30 wt% of the bio-ink composition,
Bio-ink composition.
According to claim 1,
The antibiotic is a substance, penicillin (Penicillin), nafcillin (Nafcillin), ampicillin / sulbactam (Ampicillin / sulbactam), amoxicillin (Amoxicillin), cefazolin (Cefazolin), cephalexin (Cephalexin), cephradine (Cephradine) ), Cefadroxil, Clindamycin, Doxycycline, Trimethoprim/sulfamethoxazole, Vancomycin, Linezolid, Bacitracin , which comprises at least one selected from the group consisting of gentamicin, fusidic acid and mupirocin,
Bio-ink composition.
According to claim 1,
Wherein the antibiotic component is 0.00000001 wt% to 30 wt% of the bio-ink composition,
Bio-ink composition.
According to claim 1,
The bio-ink composition further comprises a crosslinking agent;
Bio-ink composition.
9. The method of claim 8,
The crosslinking agent is, tannic acid, genipin, transglutaminase ((Moo Glo TI) Transglutaminase), ethyldimethylaminopropylcarbodiimide [1-ethyl-3(3-dimethyl aminopropyl) carbodiimide; EDC], hydroxysuccinimide (N-hydroxysuccinimide; NHS), dimethylaminopropyl ethylcarbodiimide hydrochloride (EDAC), hydroxybenzotriazole (hydroxybenzotriazole) and glutaraldehyde (Glutaraldehyde) consisting of Which comprises at least any one selected from the group,
Bio-ink composition.
9. The method of claim 8,
The cross-linking agent, 0.00001 wt% to 20 wt% of the bio-ink composition,
Bio-ink composition.
Preparing a bio-ink composition according to any one of claims 1 to 10;
outputting the bio-ink composition as a three-dimensional structure using 3D printing; and
curing the three-dimensional structure by adding a crosslinking agent;
containing,
A method for manufacturing a three-dimensional scaffold.
12. The method of claim 11,
The step of preparing the bio-ink composition,
preparing a biodegradable material;
dialysis of the biodegradable material;
centrifuging the dialyzed biodegradable material; and
mixing a growth factor material and an antibiotic component with the centrifuged biodegradable material;
which includes,
A method for manufacturing a three-dimensional scaffold.
12. The method of claim 11,
The step of outputting a three-dimensional structure using 3D printing of the bio-ink composition,
outputting the bio-ink composition in a cooling jacket at 0 °C to 20 °C; and
cooling on a 0 °C to -100 °C cooling stage;
which includes,
A method for manufacturing a three-dimensional scaffold.
12. The method of claim 11,
The crosslinking agent is, tannic acid, genipin, transglutaminase ((Moo Glo TI) Transglutaminase), ethyldimethylaminopropylcarbodiimide [1-ethyl-3 (3-dimethyl aminopropyl) carbodiimide; EDC], hydroxysuccinimide (N-hydroxysuccinimide; NHS), dimethylaminopropyl ethylcarbodiimide hydrochloride (EDAC), hydroxybenzotriazole (hydroxybenzotriazole) and glutaraldehyde (Glutaraldehyde) consisting of Which comprises at least any one selected from the group,
A method for manufacturing a three-dimensional scaffold.
A three-dimensional scaffold prepared by the method of claim 11 .
16. The method of claim 15,
The three-dimensional scaffold is,
a compressive modulus of 0.00001 MPa to 10 MPa;
Surface roughness (Ra) is 10 nm to 10 ㎛,
That the water-absorbability (water-absorbability) is 0.1% or more,
3D scaffold.

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