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WO2017192004A1 - Procédé d'alimentation en encre pour impression 3d et procédé d'impression 3d l'utilisant - Google Patents

Procédé d'alimentation en encre pour impression 3d et procédé d'impression 3d l'utilisant Download PDF

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Publication number
WO2017192004A1
WO2017192004A1 PCT/KR2017/004687 KR2017004687W WO2017192004A1 WO 2017192004 A1 WO2017192004 A1 WO 2017192004A1 KR 2017004687 W KR2017004687 W KR 2017004687W WO 2017192004 A1 WO2017192004 A1 WO 2017192004A1
Authority
WO
WIPO (PCT)
Prior art keywords
ink
dimensional printing
printing
cross
dimensional
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/KR2017/004687
Other languages
English (en)
Korean (ko)
Inventor
안근선
진송완
심진형
윤원수
강동구
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Industry Academic Cooperation Foundation of Korea Polytechnic University
T&R Biofab Co Ltd
Original Assignee
Industry Academic Cooperation Foundation of Korea Polytechnic University
T&R Biofab Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Industry Academic Cooperation Foundation of Korea Polytechnic University, T&R Biofab Co Ltd filed Critical Industry Academic Cooperation Foundation of Korea Polytechnic University
Priority to JP2018557912A priority Critical patent/JP6898353B2/ja
Priority to US16/097,863 priority patent/US11534527B2/en
Priority to EP17792906.4A priority patent/EP3453523A4/fr
Priority to CN201780032352.7A priority patent/CN109153182B/zh
Priority claimed from KR1020170056273A external-priority patent/KR20170124972A/ko
Publication of WO2017192004A1 publication Critical patent/WO2017192004A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/20—Polysaccharides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/14—Macromolecular materials
    • A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
    • A61L27/24—Collagen
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
    • A61L27/38—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix containing added animal cells
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C67/00—Shaping techniques not covered by groups B29C39/00 - B29C65/00, B29C70/00 or B29C73/00

Definitions

  • the present invention relates to a method for supplying ink to an ink receiving portion of a three-dimensional printing apparatus, a three-dimensional printing method including the method, and a three-dimensional printing apparatus to which the method is applied, and more particularly, to an ink receiving portion provided in the three-dimensional printing apparatus.
  • the present invention relates to a method of providing ink by a three-dimensional printing method and a three-dimensional printing apparatus to which the method is applied. Using the method according to the invention, it is possible to print a highly structured biological tissue shape with high precision.
  • Regenerative medicine and tissue engineering mainly use the method of making three-dimensional tissue structures as a field for repairing or replacing damaged organs, and extrusion printing based bioprinting is one of the most suitable methods for making such three-dimensional tissue structures.
  • Head models used in bioprinting technology are largely divided into ink jet-based printing and extrusion-based printing.
  • various methods using laser, ultrasound, and the like have been proposed, but the first two models are most widely used.
  • the physical properties of the bio inks required by such bio printing techniques vary greatly depending on the printing head models used. Discharge-based printing systems that use fine nozzles and syringes are more effective than jet-based technologies. Viscosity is not limited. As a result, the range of applicable biomaterials becomes much wider than that of inkjet-based printing technology. In addition, it is easy to process the thick layer, it is easy to manufacture the cell structure of the size required by the clinical.
  • the resolution of bioprinting techniques based on extrusion laminated molding developed to date is hundreds of micrometers, but the basic structure of tissues and organs in the human body is tens of micrometers or less. In particular, the diameter of the capillaries that supply nutrients to cells constituting organs or tissues is 3 to 4 micrometers, so it is difficult to implement the current bioprinting technique.
  • multiple inks may be used for bioprinting.
  • the hydrogel and the curing agent are cured together, or when co-culture of several kinds of cells, the function of the cells is improved when the division between heterogeneous cells is used rather than the method of simply mixing and spraying the cells.
  • bioprinting should be performed by spraying multiple inks. According to this conventional method, this problem has been solved by using multiple heads containing heterogeneous materials, but the printing process time increases, which adversely affects cell viability and complicates the system. In order to increase the resolution of the printing technique, a nozzle with a small diameter should be used.
  • At least one ink is supplied to an ink receiving portion by a three-dimensional printing method and three-dimensional by using one ejection, in order to produce a highly precise biological tissue shape of a complex structure using a three-dimensional printing method.
  • the present invention relates to a printing apparatus for producing a printed matter having a three-dimensional pattern by using one ejection containing two or more different inks and a printing method using the same.
  • An object of the present invention is to provide an apparatus for printing a biological tissue shape having a complex structure with high precision and resolution, and a printing method using the same.
  • the present invention is to provide a printing apparatus and printing method that can significantly reduce the shear stress of the cell while heterogeneously printing the desired shape.
  • the present invention relates to a three-dimensional printing method comprising an ink providing step of providing at least one type of ink as an ink printed matter printed by a three-dimensional printing method to a three-dimensional printing ink ejecting member including an ink receiving portion and an ejecting portion.
  • the printing method may be a bioprinting apparatus and method used for manufacturing artificial tissues, organs, and the like.
  • the step of providing at least one type of ink in the space of the ink discharge member including the ink containing portion and the discharge portion, provided in the three-dimensional printing apparatus is applied by applying a physical force to the received ink Discharging ink to a single discharge port to produce an ink discharge, and printing the ink discharge on a substrate.
  • the providing of the ink may include providing at least one ink as an ink substrate printed by a three-dimensional printing method, and when using two or more different inks, another at least one ink may be used in addition to the three-dimensional printing method. It may be an ink layer provided by the providing method. Specifically, the ink filling and the at least one second ink filling the at least one first ink in the ink receiving portion It can be provided as an ink printed matter which is printed and injected by a three-dimensional printing method.
  • the ink printed matter provided to the ink receiving member in the ink providing step may itself have a two-dimensional or three-dimensional pattern, and a fragment of the ink receiving member may also have a pattern. It is particularly useful when the printed material is an artificial organ, because it consists of one or more structures of various materials.
  • the ink ejecting member may include an ink filling and an ink ejection to have a cross-sectional pattern of the same shape as the three-dimensional printed matter. Applying a physical force to the ink contained in the space, discharging ink through a discharge portion and a nozzle connected thereto to produce an ink discharge product having a cross-sectional pattern having the same shape as a print, and printing the ink discharge onto a substrate. It relates to a three-dimensional printing method of a printed matter having a cross-sectional pattern, comprising the step of.
  • the discharge may have a cross-sectional pattern of the same shape as the final printed matter
  • the ratio can be reduced to a ratio of 100: 99 to 100: 0.1, or 100: 50 to 100: 1, 100: 18 to 100: 1.
  • the cross-sectional diameter ratio of the primary ink prints provided by the three-dimensional printing method, or the cross-sectional area ratio, or the cross-sectional diameter ratio of the secondary prints of the primary ink prints may be 100: 99 to 100: 0.1. Or, it may be reduced in a ratio of 100: 50 to 100: 1 and 100: 18 to 100: 1.
  • Another example of the present invention is a three-dimensional printing apparatus of a printed matter having a cross-sectional pattern, and includes an accommodating part for receiving ink for printing in an inner space, and having a single passage positioned below the accommodating part and passing the ink therein and accommodated in the accommodating part.
  • An ink ejecting member comprising an ejecting portion for ejecting ink,
  • the printing apparatus may include a means for applying a physical force to the received ink, and for example, may apply a force using a pressure device or a screw. In order to apply the pressure under the same conditions, The pressure may be performed using a single pressing member, or may be performed by pressing the same pressure using two or more pressing members.
  • the ink contained by applying a physical force is discharged to the discharge portion to produce an ink discharge, and printing the ink discharge on the substrate.
  • the three-dimensional printing method according to the invention it can be produced a printed matter having a cross-sectional pattern.
  • At least one ink may be further provided as an ink layered product by a method other than a three-dimensional printing method.
  • the ink laminated material is provided by the layered method rather than the three-dimensional printing method.
  • the layering method by a method other than the three-dimensional printing method there is a method of layering using a tube or a syringe and the like, and is not particularly limited. Applying the physical force is Im, and a means for pushing the ink supplied to the ink receiving member as part ejection can be performed using a ", can be used, for example the pressure member and the screw or the like.
  • applying the pressure may be performed by using a single pressing member or by using two or more pressing members.
  • the pressure may apply the pressure under the same conditions, and the pressure under the same conditions is a pressure condition under which ink can be discharged to a single discharge port to form a single ink discharge, preferably two or more different inks, or
  • the ink printed matter provided by the three-dimensional printing method and the ink layered material provided by the method other than the three-dimensional printing are ejected to a single ejection opening to form a single ink ejection. It means a pressure condition to have the same shape.
  • the three-dimensional printing method of a printed matter having a cross-sectional pattern according to the present invention can be performed using the ink ejecting member or the three-dimensional printing apparatus including the ink ejecting member according to the present invention.
  • a three-dimensional printing apparatus of a printed matter having a cross-sectional pattern includes an accommodation part accommodating ink so as to have a cross-sectional pattern having the same shape as a printed matter, and a single passage located below the accommodation part. And an ink discharge member including an ejection portion for ejecting ink contained in the accommodation portion, and further comprising a nozzle connected to the discharge portion, and a pressurizing member for applying pressure to the ink contained in the partitioned space.
  • an accommodation part accommodating ink so as to have a cross-sectional pattern having the same shape as a printed matter, and a single passage located below the accommodation part.
  • an ink discharge member including an ejection portion for ejecting ink contained in the accommodation portion, and further comprising a nozzle connected to the discharge portion, and a pressurizing member for applying pressure to the ink contained in the partitioned space.
  • the three-dimensional printing apparatus is a substrate for printing the discharge
  • Additional devices for printing and printing may include components included in conventional three-dimensional printing devices.
  • three-dimensional of a print having a cross-sectional pattern A printing method, providing an ink of an accommodating part of the ink ejecting member, applying pressure to the ink contained in each partitioned space, ejecting ink through the ejecting part, and forming an ink ejection having a cross-sectional pattern having the same shape as a printed matter. Manufacturing, and printing the ink discharge on a substrate.
  • at least one ink may be provided as an ink printed matter printed by a three-dimensional printing method, and at least one ink may be further provided as an ink layered product by a method other than the three-dimensional printing method.
  • the method of manufacturing a printed matter having a cross-sectional pattern using an ink ejecting member or a three-dimensional printing apparatus including the ink ejecting member, and a three-dimensional printing apparatus including the ink ejecting member or the ink ejecting member is a component of the apparatus.
  • the steps of the star and method will be described in detail below.
  • An ink ejecting member has a receiving portion in which ink is received, and a single passage positioned below the receiving portion so as to have a cross-sectional pattern having the same shape as a printed matter, and having a single passage through which the ink passes, and ejecting the ink contained in the receiving portion. And a discharge part.
  • the present invention can print by discharging two or more different inks using one ink ejecting member, so that a single printing head can be used. Printing process time is reduced and little shear force is applied to the ink or cells contained therein. Therefore, when using the ink containing the cells using the printing method or apparatus according to the present invention, there is an advantage that the cell viability is high and the system is simple.
  • the discharge member is a component for supplying ink by pushing the ink under pressure from the outside, which is commonly used in a three-dimensional printing apparatus.
  • the pressure applied to the ink receiving member may vary depending on the concentration of the ink and the size of the nozzle, for example, 0.1 to 700 kPa, 1 to 500 kPa or 1 to 700 kPa. have. If the printing pneumatic is high, too much material is discharged, and if the pneumatic pressure is low, subsequent material may not print and slip into the supporting material.
  • the ink ejecting member has a single passage located below the accommodating portion and has a single passage through which the multiple inks pass, and includes an ejecting portion for ejecting the ink contained in the accommodating portion.
  • the discharge part may have a single passage, and may discharge ink contained in the accommodation part divided into a plurality of spaces by a single channel control method instead of multi channel control.
  • the inner diameter of the discharge port is formed very small.
  • the ink contained in the accommodation portion through the discharge port can be discharged to the outside of the accommodation portion.
  • the ink discharge discharged from the discharge portion has a cross-sectional pattern that is the same as the cross-sectional pattern of the accommodation portion, but may be reduced in size.
  • the three-dimensional printing apparatus may further include a nozzle connected to the discharge end of the discharge member, the ink is discharged through the exposure, the plate is located under the nozzle is discharged from the ' Printing ink is laminated on the plate to produce printed matter.
  • the printed matter having a cross-sectional pattern according to the present invention is preferably a human simulated tissue.
  • a human simulated tissue For example, muscle tissue (bundle structure), bone tissue (lamel lae & canal structure), neural tissue (per ineurium structure), vascular tissue (mul t i_layer structure), spinal cord (spinal cord) tissue and the like.
  • a housing having the same cross-sectional pattern and a discharge part for discharging the same into a single passage may be provided to realize high resolution.
  • the method for printing a living tissue provides a printing ink having an enlarged cross-sectional shape having the same shape and proportion as a printed matter in the accommodating portion, while maintaining a same cross-sectional pattern of the accommodating portion,
  • the pressure can be controlled to pass through the discharge section having a small cross section. It is preferable that the cross-sectional pattern of the discharged ejection is at a level capable of maintaining the same shape as the cross-sectional pattern of the accommodation portion.
  • the meaning of 'same' is defined as a meaning including not only 100% identical but also identical enough to perform substantially the same function.
  • the cross-section maintains the same shape means that only the size of the cross section is reduced but the shape of the original cross-section remains intact. After fabrication, printing of the desired size, ie the actual tissue cells and the tissue cells that correspond to the size, is possible.
  • the viscosity of the printing ink is preferably such that the cross section of the discharged discharged through the nozzle can maintain the same shape as the cross section of the printing material.
  • the cross-section of the printed result is a small size that cannot be achieved with current bioprinting techniques or a microstructure that can significantly reduce survival when printed.
  • muscle tissue (bundle structure), bone tissue (lamel) lae & canal structure), nerve tissue (per ineurium structure), vascular tissue (mul ti-layer structure), spinal cord (spinal cord) tissue, and the like.
  • the cross section of the printing apparatus accommodating portion has a cross-sectional pattern having the same shape as the printed matter, and the cross-sectional pattern of the accommodating portion is the discharge or
  • the ratio of the cross-sectional pattern of the printed matter may be represented by various methods such as the area ratio of the cross section, the diameter ratio, and the like.
  • the cross-sectional diameter ratio of the primary ink prints provided by the three-dimensional printing method is 100: 99 to 100: 0.1. Or 100: 50 to 100: 1, 100: 18 to 100: 1 ratio.
  • the reduction ratio of the secondary prints is that the ink prints provided on the ink receiving member are subjected to two printing processes, and when the reduction ratio is expressed based on the ink prints printed first in the syringe, the syringes are three-dimensional printing.
  • Equation 1 is obtained by subtracting the diameter of the secondary printed matter or ejection of the printing ink ejected through the nozzle from the diameter of the primary ink printed matter provided by the ratio divided by the diameter of the primary ink printed matter provided to the syringe by the three-dimensional printing method. It can be expressed as
  • A is the diameter of the primary ink substrate provided in the syringe by three-dimensional printing
  • B is the diameter of the secondary print of the printing ink
  • a and B are the same length unit.
  • the reduction ratio of the secondary printed matter is directly affected by the cross-sectional diameter of the receiving portion, the fragment diameter of the discharge portion, or the diameter of the nozzle, and can be variously designed by appropriately adjusting to the cross-sectional pattern size of the desired printed matter.
  • the cross-sectional diameter of the printout varies depending on the size of the nozzle, which ranges from 0.1 mm to lmm, and is usually used for printing, such as material properties, pressure, speed of the printing head, and the location of the printing result (printing bed). According to the process Can change.
  • the ratio can be downsized to 99% or 98.7% (200 ⁇ ) from the total diameter (15 ⁇ s) of a particular shape (eg Lobule).
  • the ink provided to the ink ejecting member according to the present invention is preferably a bio ink capable of producing artificial organs or the like.
  • printing may be performed by providing ink to the ink containing portion so as to have a cross-sectional pattern having the same shape as the printed matter.
  • the different inks mean that the inks are different from one or more selected from the group consisting of constituents, constituent contents and physical properties.
  • the term 'bio ink' includes living cells or biomolecules, and is a term used to collectively refer to a material capable of fabricating a structure required for a bioprinting technology.
  • the bio ink of the present invention comprises a liquid, semisolid, or solid composition comprising a plurality of cells.
  • the bio ink should provide a physical environment for three-dimensional processing and a biological environment for the cells to perform the intended function.
  • the supply of nutrients and oxygen necessary for the survival of cells in the ink ejecting member is appropriately performed. It should also be able to protect the cells from the physical stresses that occur during the printing process.
  • the bio ink should have the physical properties required in the printing process such as repeatability of 3D patterning, productivity, and no clogging of the nozzle.
  • the ink according to the present invention is preferably a hydrogel, and thus
  • the ink may include a gelling polymer and may include, for example, one or more selected from the group consisting of gelling polymers, cells, growth factors, and extracellular substrates.
  • the bioink used in the present invention may be, for example, a mixed / non-mixed hydrogel of desired cells, a hydrogel containing a specific growth factor, a hydrogel containing a cell and a growth factor, a cytokine (cytokine). ) Containing hydrogels, different types of hydrogels.
  • Hydrogels include collagen, matrigel, alginate, gelatin and agarose. Decellularized tissue-derived cell ink, hyaluronic acid, fibrin gel, or the like or a compatible hydrogel is suitable.
  • bio-inks diffuse faster at lower viscosities, they are thicker than water (1 cp) and have a viscosity measured at 25 ° C, ranging from 2 cp to 1,000,000 cp, for example 2 cp to 10,000 cp, 5 cp to 1,000,000 cp, 2 to Materials on gels having a viscosity of 500 cps, 5 to 300 cps and the like are suitable.
  • the viscosity of the ink is too low, the shape of the ink printed matter printed by the three-dimensional printing method may collapse or be deformed, and the printed matter and the ink packing may be mixed with the boundary to be broken.
  • the viscosity of the gel-type material used in the method of the present invention preferably has a suitable viscosity so that the printing material can be discharged in the discharge process described later.
  • the inks applicable to the present invention may use various viscosity enhancers to provide a viscosity suitable for ejection. Viscosity of the printing material is such that the cross section of the discharged discharged through the nozzle can maintain the same shape as the cross section of the printing material.
  • the ink viscosity difference between the ink layer and the ink printed matter is less than 5,000cp, for example 0 to 5,000cp, less than 1,000cp, less than 500cp, less than 200cp, less than 150cp, less than lOOcp It may be less than 50cp. If the difference in viscosity of the ink is very large, the shape of the ink printed matter may be deformed by the molecular force of different materials, the same if you want to three-dimensional printing by ejecting When pressure is applied to the ink or the ink receiving member, the ink pattern accommodated in the ink receiving member may collapse due to the difference in viscosity. Therefore, different from each other
  • the difference between the elastic values of the ink filling and the ink printing may be 10,000 Pa or less, for example, 0 to 10,000 Pa. It is preferable that the ink layered matter and the ink printed matter have similar trends in viscosity and elasticity change according to the shear rate and similar viscosity and elasticity values.
  • the gelling polymer used in the different inks is different, for example, the gelling polymer is temperature sensitive like collagen and gelatin, and the other is not temperature sensitive like alginate and fibrin gel, it is necessary to adjust the temperature inside the syringe.
  • the temperature of the ink accommodating member can be appropriately adjusted within the 4 S C to 37 C C temperature range.
  • Naturally derived or synthetic hydrogels in the field of three-dimensional bio printing Although bio inks have been developed and are currently in use, biogels based on hydrogels have been used for physical and biological reasons such as biocompatibility, printing suitability, geometrical precision, and precision.
  • the first extrudable "or" ejection possible ' is the discharge portion, the nozzle (nozzle) or an orifice (for example, one or more holes or tubes) means that can be molded by passing (e.g., under pressure).
  • Densification of the bio ink is derived from growing cells at a suitable density. The cell density required for the bio ink depends on the cell to be used and the tissue or organ to be produced.
  • the present invention also provides a bio ink composition, wherein the bio ink composition further comprises a tissue-derived component.
  • Tissue-derived component means that specific tissues of animals such as cartilage, kidney, heart, liver, muscle, etc. are decellularized and gelled of a substance mainly composed of extracellular matrix. Can be included to enhance.
  • the bio ink composition is a cell culture medium It may further include.
  • the cell culture medium is a concept including any medium suitable for the cells of interest.
  • the ink according to the present invention may include a gelling polymer, and the gelling polymer solution used for such printing may be used in various kinds.
  • the conditions that the polymer solution should have are as follows. First, in order to achieve a three-dimensional printing well, it should be easy to eject to the nozzle with a suitable viscosity, and should not cause problems such as crushing the shape of the object made by rapid curing after discharge. In addition, for manufacturing purposes, it should be possible to create a cell culture environment similar to the tissue in the human body.
  • gelling polymer examples include fucoidan, collagen alginate, chitosan, hyaluronic acid, silk, polyimides, polyamix acid, polycarprolactone, polyetherimide, nylon (nylon) , Polyaramid, polyvinyl alcohol, polyvinylpyrrolidone,
  • Polyaniline polyacrylonitrile, polyethylene oxide, polystyrene, cellulose, polyacrylate, polymethylmethacrylate ), Polylact ic acid (PLA), polyglycol ic acid (PGA), copolymers of polylactic acid and polyglycolic acid (PLGA), poly
  • PEG-DA ⁇ Poly (ethylene oxide) terephthalate-co-butyleneterephthalate KPE0T / PBT), polyphosphoester (PPE), polyphosphazene (PPA), polyanhydride (PA), polyortho Ester ⁇ poly (ortho ester; P0E), poly (propylene fumarate) -diacrylate; PPF-DA ⁇ and polyethylene glycol diacrylates ⁇ poly (ethylene glycol) diacrylate; Out of the group consisting of PEG-DA ⁇ It may be one or more selected or a combination of the above materials. However, the material is not limited to this embodiment.
  • the gelling polymer may be a chemically modified natural polymer, for example, the binding si of GelMA, Alginate / Gel at in, Alginate that combines the chemical at the gel at in and methacrylate (MA) and the photoini ti ator
  • the stock price may include pentapeptide sequencing, Tyr-I le-Gly-Ser-Arg (YIGSR) and alginate in combination with EDC / NHS.
  • the hydrogels including polyethylene glycol, alginate, nitrate, collagen and gelatin have high water content, have excellent biocompatibility, can control mechanical properties, and have excellent biodegradability, and thus are widely used in the preparation of cellular-embedded carriers. Has been used. For this reason, hydrogels are well suited for the manufacture of cell-loaded structures and can be printed directly to obtain various types of tissue regeneration backbones.
  • the gelatin is particularly suitable as the cell carrier because it exhibits temperature-sensitive properties. That is, gelatin has the property of liquefying at 37 ° C and solidifying below room temperature.
  • the gelling polymer may be formed using a physical treatment or a chemical treatment to form a crosslinking ol
  • the crosslinking solution may be used for the chemical treatment
  • a crosslinking solution may be appropriately selected according to the selected gelling polymer.
  • the crosslinking solution may be gypsum; Or hydroxyapatite, apatite carbonate, apatite fluoride, apatite chloride, a-TCP, ⁇ -TCP, calcium metaphosphate, tetracalcium phosphate, calcium hydrogen phosphate, calcium dihydrogen phosphate, calcium pyrophosphate, calcium carbonate, calcium sulfate, EDC ⁇ l-ethyl- (3-3-dimethylaminopropyl) carbodi imide hydrochlor ide ⁇ or a solution of one or more combinations thereof selected from salts thereof.
  • the ratio of collagen concentration in the collagen solution of the liquid form is usually in the range of 0.01 to 30% by weight.
  • the method for producing hydrogels uses ink for normal three-dimensional printing. It can be carried out by applying the manufacturing method used when manufacturing, but is not particularly limited,
  • the bioink according to the present invention may include cells, and applicable cells or tissues are not particularly limited, and may be animal cells or plant cells, or tissues of animals or plants.
  • the cells are stem cells, osteoblasts, myoblasts, myocytes, tenocytes, neuroblasts, fibroblasts, glioblasts, germ cells ( germcells, hepatocytes, renal cells, sertoli cells, chondrocytes, epithelial cells, cardiovascular cells, keratinocytes, smooth muscle cells cells, cardiomyocytes, glial cells, endothelial cells, hormone secreting cells, cotton cells, pancreatic islet cells and neurons. It may be any one or more.
  • the cell type used in the prepared artificial tissue of the present invention may be cultured in any manner known in the art. Cell and tissue culture methods are known in the art.
  • the cells may also be incubated with cell differentiation materials that induce differentiation of the cells along the desired cell line.
  • stem cells are incubated in contact with differentiation medium to produce a range of cell types.
  • differentiation media include, but are not limited to, osteogenic differentiation medium, chondrogenic differentiation medium, adipogenic differentiation medium, neuronal differentiation medium, cardiomyocyte differentiation medium, and enterocyte differentiation medium (e.g., intestine). Incubated in contact with the differentiation medium).
  • the cells may be cultured with growth factors, cytokines and the like.
  • Growth factor refers to a protein, polypeptide, or polypeptide complex comprising a cytokine, produced by a cell and capable of affecting itself and / or various other adjacent or isolated cells.
  • the growth factor is In response to developmental or multiple biochemical or environmental stimuli, it affects the growth and / or differentiation of certain types of cells. Some, but not all, growth factors are hormones.
  • Exemplary growth factors include fibers including insulin, insulin-like growth factor (IGF), nerve growth factor (NGF), vascular endothelial growth factor (VEGF), keratinocyte growth factor (KGF), and basic FGF (bFGF) Platelet-derived growth factor (PDGF) including blast growth factor (FGF), PDGFM and PDGF-AB, bone morphogenic proteins (BMP), hepatocyte growth factor (HGF), traits including BMP-2 and ⁇ P-7 Transforming growth factor beta (TGF- ⁇ ), epidermal growth factor (EGF), granulocyte-macrophage colony-stimulating factor (GM) including transforming growth factor alpha (TGF- ⁇ ), TGF
  • IGF insulin-like growth factor
  • NGF nerve growth factor
  • VEGF vascular endothelial growth factor
  • KGF ker
  • 'bioprinting' refers to three-dimensional accurate cell deposition (eg, cell solutions, cell-containing gels, cell suspensions, cells) through a methodology compatible with automated computer-assisted, three-dimensional prototyping devices (eg bioprinters). Concentrate, multicellular aggregates, multicellular bodies, etc.). 3D printing can be performed by extruding a biodegradable polymer from a nozzle using a bio-plotter and laminating it on a stage.
  • tissue analogues can be generated by the methods described above.
  • the pattern or stacking arrangement for stacking the bio ink composition may be determined by the size and diameter of the tissue-like organ to be manufactured.
  • the number of cells included in the bio ink used to manufacture tissue-like organs may be controlled according to the type of cells, the content of cell nutrients contained in the bio ink composition, and the like.
  • the type of cells included in the bio ink composition may be variously changed according to the type of tissue-like organ to be prepared according to the above method. Those skilled in the art to which the present invention pertains will be able to select and apply appropriate cells according to the type of tissue-like organs to be prepared through three-dimensional bioprinting.
  • Bio ink composition is sprayed by a three-dimensional bio printer laminated Thereafter, it is possible to promote crosslinking of the bio ink ⁇ composition by heating it, exposing it to ultraviolet rays or adding a crosslinking solution. This crosslinking allows the laminated bio ink composition to be completed into a more rigid structure.
  • An optical initiator may be used to promote the crosslinking.
  • an ink ejecting member having the same cross-sectional pattern as the printed matter but having the same or different cross-sectional pattern
  • at least one ink provided by a three-dimensional printing method is accommodated in a space of the ink ejecting member accommodating portion, It is provided with a single passage through which the ink passes and controls the pressure, for example the piston, to pass through the discharge portion for discharging the ink.
  • a small amount of hydrogel as a support material in the amount of 0.01 mL to 2 mL in advance so as not to pour the bio ink into the container. Thereafter, or without adding hydrogel to the barrel.
  • the filled hydrogel is ejected at the beginning of printing, and then the desired shape is printed. After printing the printed material of the desired shape, the hydrogel filled above will come out.
  • the reason for adding support material (support mater al) at the beginning and the end is to make printing stable.
  • 1, 2, 3, 4, and 5 schematically illustrate a process of providing ink to a ink receiving member by a three-dimensional printing method according to an example of the present invention.
  • the inside of the ink discharge member 10 is divided into a plurality of spaces for accommodating the respective inks 11, 12, 13, and 14.
  • 9 shows a bioprinting apparatus which can be applied to an example of discharging five inks (1, 2, 3, 4, 5) according to one embodiment of the present invention.
  • the cell liquid substance contained in the ink ejecting member 10 is pressed in the A direction and printed through the ejecting portion 20 or the nozzle 80.
  • the ink is ejected to complete the final object 50.
  • the piston 60 is preferably controlled to pass through the nozzle 20 is reduced in size while maintaining the same shape of the cross section of the printing material, the base may be a container containing a liquid material in some cases.
  • the pressure is too high, the load on the nozzle may be increased and damage may occur, or the hydrogel may not be discharged in the form of a thread smoothly and may be discharged in an unbalanced shape, and the pressure may be too weak. In this case, due to the resistance due to the viscosity of the hydrogel may not be smooth discharge from the nozzle.
  • the diameter is too small, the discharge pressure is increased, so that the risks when the pressure is strong may occur the same, and if the diameter is too large, the precision of the three-dimensional shape when manufacturing the scaffold may be reduced.
  • the printing method according to the present invention includes the steps of accommodating ink in the accommodating part, applying a pressure within the range of 0.1 to 700 kPa, 1 to 500 kPa or 1 to 700 kPa, for example, 0. Discharging the ink to the nozzle having an outlet diameter within a range of 1 to 1 mm, and printing the ink while the nozzle moves at a speed within a range of 1 to 700 mm / min by a moving part of a printing apparatus. Can be done.
  • the cell liquid substance contained in the ink discharge member 10 is pressed in the A direction to discharge the printing material to the base 100 through the discharge portion 20 to complete the final object 50.
  • the piston is preferably controlled to pass through the discharge portion 20 is reduced in size only while maintaining the same shape of the cross section of the printing material, in some cases the base may be a container containing a liquid material.
  • the base may be a container containing a liquid material.
  • the volume reduction of the material because there is a limit to the size reduction of the nozzle inner diameter.
  • the volume of material discharged in proportion to the number of multiple inks can be reduced, so that precise jetting is possible compared to the prior art.
  • the area of contact between each material and the inner surface of the nozzle passage is reduced, and the shear force generated is also reduced than when discharging a single material. Therefore, in terms of improving cell activity and printing accuracy, there is an advantageous effect compared to the prior art.
  • FIG. 1 is a schematic diagram illustrating a process of manufacturing a printed matter of a second ink and a layered ink of a first ink according to an embodiment of the present invention.
  • FIG. 2 is a schematic view showing an ink ejecting member into which a first ink layered product and a spiral second ink printed matter are injected and an ejection obtained therefrom according to an example of the present invention.
  • FIG. 3 is a schematic view showing an ink ejecting member into which a print made of three kinds of ink of crab 1 ink and a spiral shape is injected, and an ejection obtained therefrom according to an example of the present invention.
  • FIG. 4 is ink ejected by a three-dimensional printing method according to an embodiment of the present invention A diagram schematically showing an example of injecting ink into a member.
  • FIG. 5 is a view schematically illustrating a method of injecting the second to seventh inks by a three-dimensional printing method in addition to the filling of the first ink in the ink ejection member according to an embodiment of the present invention.
  • FIG. 6 is a photograph of a syringe and a discharging part provided at the end of the syringe showing a process of manufacturing a print of the crab 1 ink and the crab 2 ink according to Example 1
  • FIG. 7 is a confocal micrograph showing the result of printing using the ejection obtained from the ink ejecting member in which the filling of the first ink and the printed matter of the spiral second ink were injected according to Example 1.
  • FIG. 7 is a confocal micrograph showing the result of printing using the ejection obtained from the ink ejecting member in which the filling of the first ink and the printed matter of the spiral second ink were injected according to Example 1.
  • FIG. 8 is a confocal micrograph showing the result of printing using a syringe in which a layered material of crab 1 ink and a printed material of a second ink are laminated according to Example 2, and an ejection obtained from the syringe.
  • FIG. 9 is a view schematically showing an ink ejecting member having a receiving portion in which five different inks are layered according to an example of the present invention.
  • 10 is a view schematically showing an ink ejecting member according to an embodiment of the present invention.
  • Example 1 A syringe including an ink substrate and an ink layer
  • a 1-gel hydrogel for layering of 3 w / v% sodium alginate was injected into a syringe, which is an ink receiving portion of a 3D printing apparatus.
  • a 3D printing apparatus having a long length nozzle was used.
  • 3 w / v% sodium alginate containing green fluorescent particles was injected as a 12 hydrogel by three-dimensional printing.
  • One example was performed similar to the method illustrated in FIG. 1. The ink shape printed on the syringe was photographed and shown in FIG. 2.
  • the first hydrogel for filling the syringe and the three-dimensional printed giant 12 hydrogel were printed by a three-dimensional printing method using a discharge obtained through a discharge unit by applying pressure, and the nozzle size (nozz le I. D) was 1.0 mm.
  • Printed in three lines. Printed section length is 30 micrometers when using a nozzle with a nozzle size (nozz le I .D) of 1 mm and printed section length when using a nozzle with a nozzle size (nozzl e I .D) of 2 mm Was 70 micrometers.
  • FIG. 3 shows a photograph in which a first hydrogel for supporting and a second hydrogel having a specific shape formed therein by a three-dimensional printing method are formed using a discharge member including an ink receiving unit according to Example 1.
  • FIG. 7 shows the results of confocal microscopy observation of a printed matter produced by a three-dimensional printing method by ejecting the ink containing layer having the first hydrogel and the crab hydrogel layered in accordance with Example 1 under pressure conditions. That is, when the discharged product is observed by fluorescence with a confocal microscope, it is as shown in FIG.
  • the method according to the invention indicates that the ink is printed with high resolution.
  • Example 2 three-dimensional printing using nozzles of various sizes
  • Example 2 Using a three-dimensional printing apparatus using the same ink ejecting member as Example 1, it was confirmed by confocal microscopy that the hydrogel is printed in the nozzle size of 18, 20, 22, 25 and 27 Gauge. Specifically, the nozzle inner diameters of 18, 20, 22, 25, and 27 Gauge were 0.82 mm, 0.63 mm, 0.41 mm, 0.28 mm, and 0.1 mm. The confocal micrograph is shown in FIG. 8.
  • FIG. 8 shows the first hydrogel layered product and the second hydrogel printed matter of the size of the nozzle using the discharge member including the ink containing portion according to Example 2.
  • A is the diameter of the primary ink substrate provided in the syringe by three-dimensional printing method B is the diameter of the secondary substrate of the printing ink
  • a and B are the same length unit.
  • hydrogel was injected into the same syringe as in Example 1.
  • the hydrogel is injected, the syringe, using the future a three-dimensional printing, comprising a long length of the nozzle, the ink printed print each Green, Blue, Red fluorescent particle is a 3 w / v% sodium alginate comprising sequentially It was injected into a syringe, and it was confirmed by confocal microscopy that the obtained RGB hydrogel was printed.
  • FIG. 7 A schematic diagram of a syringe filled with ink prints prepared with the three inks and a manufacturing process thereof are shown in FIG. 7.
  • 3 w / v% sodium alginate was injected into the same syringe as in Example 1, and 3% gelatin, a temperature-sensitive hydrogel, was injected into the alginate pre-injected using a long nozzle using a printing method.
  • Printing the prepared complex hydrogel in 200 Mm calcium chloride induces only alginate gelation and gelatin is present in the ungelled body.
  • the printing structure was placed in a liquid of 37 a C, the gelled alginate remained intact but gelatin melted to form a lumen structure.
  • Example 5 Blood Vessel Fabrication with Cells and Multiple Lumen Structures
  • 3 w / v% sodium alginate was injected into the syringe and smooth muscle was used, using a three-dimensional printing apparatus equipped with a long nozzle.
  • 3% alginate in which the cells are contained at a concentration of at least IX 10 7 Cel ls / mL is injected into the filled 3% alginate, and 3% gelatin embedded at a concentration of at least 1 X 10 7 Cel ls / mL is used for smooth muscle cells.
  • Vascular structures were simulated by a continuous method of injecting the intrinsic alginate.
  • the aorta and the vena cava of the blood vessels are stacked in a four-cylindrical structure. This method enables easy hand printing of quadrilateral blood vessels, as well as size control.
  • the double structure of the cleansing vein and the single structure of the microvessel can be similarly simulated.

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Abstract

La présente invention concerne un procédé pour charger deux sortes différentes, ou plus, d'encres multiples dans un élément d'émission d'encre pour l'impression 3D et un procédé d'impression 3D utilisant les encres chargées, le procédé d'impression 3D utilisant des encres multiples comprenant les étapes consistant à : produire une émission d'encre par l'émission des encres multiples contenues sous pression à partir d'un orifice d'émission unique d'une unité d'émission ; et imprimer à l'aide de l'émission d'encre.
PCT/KR2017/004687 2016-05-03 2017-05-02 Procédé d'alimentation en encre pour impression 3d et procédé d'impression 3d l'utilisant Ceased WO2017192004A1 (fr)

Priority Applications (4)

Application Number Priority Date Filing Date Title
JP2018557912A JP6898353B2 (ja) 2016-05-03 2017-05-02 三次元プリンティング用インクを供給する方法およびこれを用いた三次元印刷方法
US16/097,863 US11534527B2 (en) 2016-05-03 2017-05-02 Method for supplying inks for three-dimensional printing, and three-dimensional printing method using same
EP17792906.4A EP3453523A4 (fr) 2016-05-03 2017-05-02 Procédé d'alimentation en encre pour impression 3d et procédé d'impression 3d l'utilisant
CN201780032352.7A CN109153182B (zh) 2016-05-03 2017-05-02 用于给三维打印供应墨的方法和使用该方法的三维打印方法

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KR20160054397 2016-05-03
KR10-2016-0054397 2016-05-03
KR1020170056273A KR20170124972A (ko) 2016-05-03 2017-05-02 삼차원 프린팅용 잉크를 공급하는 방법 및 이를 이용한 삼차원 프린팅 방법
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114874461A (zh) * 2022-03-24 2022-08-09 北京大学 一种水凝胶材料及其制备方法与应用

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000280356A (ja) * 1999-03-29 2000-10-10 Minolta Co Ltd 三次元造形装置および三次元造形方法
WO2015066705A1 (fr) * 2013-11-04 2015-05-07 University Of Iowa Research Foundation Bio-imprimante et procédés pour l'utiliser
WO2015077262A1 (fr) * 2013-11-19 2015-05-28 Guill Tool & Engineering Entrées d'impression 3d coextrudées, multicouche et multicomposant
KR20160030939A (ko) * 2013-06-13 2016-03-21 애스펙트 바이오시스템즈 리미티드 3차원 구조물의 첨삭 가공을 위한 장치 및 방법
KR20160036619A (ko) * 2013-07-31 2016-04-04 오가노보, 인크. 조직을 제작하기 위한 자동화 장치, 시스템 및 방법

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000280356A (ja) * 1999-03-29 2000-10-10 Minolta Co Ltd 三次元造形装置および三次元造形方法
KR20160030939A (ko) * 2013-06-13 2016-03-21 애스펙트 바이오시스템즈 리미티드 3차원 구조물의 첨삭 가공을 위한 장치 및 방법
KR20160036619A (ko) * 2013-07-31 2016-04-04 오가노보, 인크. 조직을 제작하기 위한 자동화 장치, 시스템 및 방법
WO2015066705A1 (fr) * 2013-11-04 2015-05-07 University Of Iowa Research Foundation Bio-imprimante et procédés pour l'utiliser
WO2015077262A1 (fr) * 2013-11-19 2015-05-28 Guill Tool & Engineering Entrées d'impression 3d coextrudées, multicouche et multicomposant

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP3453523A4 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114874461A (zh) * 2022-03-24 2022-08-09 北京大学 一种水凝胶材料及其制备方法与应用

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