CN107502061B - Superficial degradation type 3D printing bio-ink and 3D printing method - Google Patents
Superficial degradation type 3D printing bio-ink and 3D printing method Download PDFInfo
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- CN107502061B CN107502061B CN201710790440.5A CN201710790440A CN107502061B CN 107502061 B CN107502061 B CN 107502061B CN 201710790440 A CN201710790440 A CN 201710790440A CN 107502061 B CN107502061 B CN 107502061B
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
- C09D11/00—Inks
- C09D11/30—Inkjet printing inks
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y10/00—Processes of additive manufacturing
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y70/00—Materials specially adapted for additive manufacturing
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09D—COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR
- C09D11/00—Inks
- C09D11/30—Inkjet printing inks
- C09D11/38—Inkjet printing inks characterised by non-macromolecular additives other than solvents, pigments or dyes
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Abstract
A kind of superficial degradation type 3D printing bio-ink and 3D printing method, the 3D printing bio-ink is made of a kind of gel ink and a kind of holder ink, the gel ink is made of hydrogel and biological cell, the hydrogel includes the PLGA PEG PLGA triblock copolymers for having temperature-responsive, and the holder ink is the makrolon lactide copolymer (PTMC PLLA copolymers) with superficial degradation characteristic.The present invention using above-mentioned 3D printing bio-ink Thermo-sensitive and can superficial degradation characteristic, the demand maintained for structure-controllable in degradation process in 3D biometric print stake bodies, utilize multi-component material gradient printing technique, the double-deck print structure of structure similar " coaxitron ", can printing shaping obtain the model for having controlled surface degradation holder support containing biological cell, and the nutrition and metabolism channel that can obtain porous structure is conducive to obtain the thicker institutional framework of bigger by 3D printing.
Description
Technical field
The present invention relates to biotechnology, especially 3D biometric prints technical field, more particularly to a kind of superficial degradation
Type 3D printing bio-ink and 3D printing method.
Background technology
3D printing can be rapidly and efficiently produce personalized product, thus be gradually introduced in biologic medical industry,
To be used for tissue and organ transplant by 3D printing technique.《The current research of biological 3D printing and application》(powder metallurgy work
Industry, the 4th phase of volume 25, in August, 2015, Zhang Hongbao etc.) in the application of 3D biometric print technologies be divided into different levels carry out
It introduces, it is incorporated herein by reference.The basic skills of the 3D biometric prints of higher level is, by seeded with living celis to biofacies
It in biodegradable holder hold and last, is then cultivated in the bioreactor so that cell relies on holder to be formed
Spatial growth generate needed for tissue.For example, 106085949 A of CN, which disclose one kind, being based on the molding reconstruction urethra of 3D printing
The method of prosthese；It is divided into epithelioid cell, class smooth muscle cell by cell inducible factors, is then converted into cell drop,
The cell drop has cell and culture solution hydrogel to mix, and the bio-ink used for medical 3D printer is made；Choosing
Select the holder ink of hydrogel that collagen is mixed with alginates as timbering material；Cell or cell aggregation are controlled by computer
Body, the eject position and dynamics of ratio and nozzle between cell and gel are lifted, 3D printing with electrical-controlled lifting platform courses nozzle
Two nozzles of machine successively printing alternate urethra section.By in vitro culture fat-derived stem cells, it is induced to be divided on urinary tract
Chrotoplast and smooth muscle cell are then converted into cell drop, in conjunction with 3D printing, rebuild urethra in vitro, then will rebuild again
Urethra be transplanted to patient with it go.Also that is, the 3D biometric print materials used in above-mentioned existing 3D biometric prints technology are two
Kind：One is celliferous bio-ink is wrapped, one is the timbering materials as holder.
The prior art is basically to find suitable bio-ink and timbering material, example about the research direction of 3D printing
Such as：104399119 A of CN disclose a kind of method preparing strong mechanical performance cartilage based on 3D biometric prints, pass through silk
Fiber and gelatin solution prepare the bio-ink containing cartilage stem cell, and forming holder with PCL materials, (PCL is a kind of semicrystalline
Polymer, it is [CH that structure, which is made, by 6-caprolactone titanium catalyst, dihydroxy or trihydroxy initiator ring-opening polymerisation2-(CH2)4-
COO]nPolyester).Contain phosphate buffer, Sodium Alginate, gelatin, methacrylic acid anhydride solution, two water wherein in bio-ink
Calcium sulfate, silk fiber and UV photoinitiators are closed, ionic bond, gelatin and metering system are formed by Sodium Alginate and calcium ion
Acid anhydrides forms the mechanical property that covalent bond improves gel, and printing, which is irradiated after completing by UV light, to be molded.For another example：CN
A kind of bio-ink for 3D printing is disclosed in 105238132 A, constituent includes water-soluble with crosslinking function
Property synthetic polymer, the water-soluble natural macromolecule with crosslinking function, the bioactivity of special ultra microstructure can be spontaneously formed
Component, cross-linked evocating agent and solvent, further comprise biological active component；Ink is finally solidified into again by the irradiation of UV light
Type.105885436 A of CN disclose a kind of bio-ink material and its preparation method and application for 3D printing, this is existing
Technology provides timbering material in fact, and the timbering material is using large biological molecule, precrosslinker, coagulant as holder ink
Material is small and with biodegradability by shape acquisition antigenicity, rejection of washing, crosslinking agent after printing
Soft tissue holder.
Described above is the background of the 3D biometric print technologies comprising holder, is disclosed in CN 103249567A
For manufacturing in the devices, systems, and methods of tissue, the component of a variety of materials of 3D biometric prints has been carried out detailed
It enumerates, wherein also whether there is or not the trials of holder 3D biometric print technologies for the specifically mentioned prior art, but exists very without support technology
More limitations, such as be difficult to obtain complicated geometry, be difficult to form the blood vessel network etc. that nutrient needed for tissue production is provided.Cause
This, what the prior art provided is the 3D biometric print solutions comprising holder.
In fact just because of above-mentioned reason, existing 3D biometric prints technology can only print a very thin layer tissue, because
To be difficult to obtain nutriment without nutrition channels, internal cells such as blood vessels compared with inside thick tissue, metabolite can not be discharged,
Thus thicker 3D printing tissue is difficult to Sustainable Growth.
Invention content
The technical problem to be solved in the present invention is to provide a kind of superficial degradation type 3D printing bio-ink and 3D printing method,
The problem of being formerly mentioned is reduced or avoided.
Specifically, the present invention provides a kind of superficial degradation type 3D printing bio-ink and 3D printing methods, for 3D
The demand that structure-controllable maintains in degradation process in biometric print stake body prepares the printable biology with superficial degradation characteristic
High molecular material can print the model for having the support of controlled degradation holder obtained containing biological cell, and can also be into
One step obtains porous model, can be produced for cell and provide nutrition and metabolism channel, be conducive to obtain bigger by 3D printing
Thicker institutional framework.
In order to solve the above technical problems, the present invention proposes a kind of superficial degradation type 3D printing bio-ink, it is solidifying by one kind
Celluloid ink water and a kind of holder ink are constituted, and the gel ink is made of hydrogel and biological cell, wherein the hydrogel by
Following component is constituted：The PLGA-PEG-PLGA triblock copolymers of mass fraction 10%~40%, mass fraction 0.001%~
1% Porcine HGF, the anti-inflammation hemostasia drug of mass fraction 0.05~0.3%, surplus are deionized water；The holder ink
Water is made of 5%~50% makrolon lactide copolymer of the gross mass of the gel ink.
Further, the present invention also provides a kind of preparation method of above-mentioned superficial degradation type 3D printing bio-ink, packets
Include following steps：By the PLGA-PEG-PLGA triblock copolymers of mass fraction 10%~40%, mass fraction 0.001%~
1% Porcine HGF, the anti-inflammation hemostasia drug of mass fraction 0.05~0.3% are blended with the deionized water of surplus, stirring
30min stands 1~6 hour and the hydrogel is made；The hydrogel and the biological cell are uniformly mixed into the 3D to beat
Print the gel ink of bio-ink.
In addition, the present invention also provides a kind of 3D printing method using above-mentioned superficial degradation type 3D printing bio-ink,
Include the following steps：
Step A：Using above-mentioned superficial degradation type 3D printing bio-ink, the gel ink is in the temperature less than phase transition
Under the conditions of, soybean protein isolate solution is added and stirs evenly, calcium sulphate soln is then added and is uniformly mixed into spare 3D printing
Gel ink material；
Step B：The temperature for controlling the gel ink sump of 3D biometric print machines is less than phase transition temperature, and printing table top temperature is
It 37 degrees Celsius, using tool there are two the 3D printer of coaxially arranged nozzle, is beaten by the inner circumferential side nozzle of the 3D printer
It prints the holder ink and forms holder, enclosed the 3D printing gel ink material by the peripheral side nozzle of the 3D printer
It is printed as model around the holder.
Preferably, in the step A, the biological cell is before preparing the 3D printing bio-ink, to the life
Object cell carries out protection processing：The biological cell and a concentration of 10% glycerine and a concentration of 10% starch solution is equal
Even mixing stands 2-3 hours.
Preferably, the above method can further include following steps：
Step C：The model is placed in temperature control box, slowly reduces the temperature in the temperature control box to 0 degree hereinafter, making institute
The moisture stated in model freezes completely；
Step D：The temperature in the temperature control box is quickly increased to 37 degrees Celsius, keeps the hydrogel in the model completely solid
Change, to obtain the porous structural model containing biological cell.
Preferably, in the step C, the temperature in the temperature control box is slowly reduced to zero with the speed of 4-5 degree per hour
Lower 15 to minus 20 degrees.
Preferably, in the step D, 37 degree of dry-heat air is conveyed into the temperature control box, makes the ice in the model
Rapid translating is escaped at gas from the model, to form porous structure on the model.
Preferably, the method further includes obtaining the porous structural model containing biological cell
Later, vascular endothelial cell is injected in the porous structure, the vascular endothelial cell is made to be attached on structure pore framework,
The model is placed in nutritive solution later and is cultivated, blood vessel, the model are gone out by the vascular endothelial cell growth
In original cell growth at required tissue, the blood vessel that grows further provides nutrient for required tissue and metabolism is logical
Road.
Preferably, the method further includes：Obtaining the porous structural model containing biological cell
Later, the parallel via holes that the model is uniformly got to perforation by laser, for the hole connection in model to be got up, then
Vascular endothelial cell is injected in the porous structure, so that the vascular endothelial cell is attached on structure pore framework, later
The model is placed in nutritive solution and is cultivated, blood vessel, the model Central Plains are gone out by the vascular endothelial cell growth
For some cell growths at required tissue, the blood vessel grown further provides nutrient and metabolic pathway for required tissue.
The superficial degradation type 3D printing bio-ink of the present invention is made of gel ink and holder ink, wherein gel ink
It is made of hydrogel and biological cell, the hydrogel includes the PLGA-PEG-PLGA triblock copolymers for having temperature-responsive
Object, the holder ink are the makrolon lactide copolymer with superficial degradation characteristic.The present invention utilizes above-mentioned 3D printing
The Thermo-sensitive of bio-ink and can superficial degradation characteristic, tieed up for structure-controllable in degradation process in 3D biometric print stake bodies
The demand held, prepare with superficial degradation characteristic printable bioabsorbable polymer material, can a printing shaping contained
Biological cell has the model of controlled degradation holder support, and can obtain the nutrition and metabolism channel of porous structure, has
The thicker institutional framework of bigger is obtained conducive to by 3D printing.
Specific implementation mode
For a clearer understanding of the technical characteristics, objects and effects of the present invention, existing through the invention specific
Embodiment is described in detail.
Just as described in the background section, no matter existing 3D biometric prints technology selects what kind of bio-ink or holder
Material, the tissue for being difficult to that printing is overcome to obtain lack the defect in nutrition and metabolism channel.Therefore, the present invention provides a kind of new
3D printing method, can by 3D printing obtain containing biological cell without holder support or one-pass molding have holder support
Model, and can also further obtain porous model, can be produced for cell and nutrition and metabolism channel be provided, be conducive to
The thicker institutional framework of bigger is obtained by 3D printing.
Specifically, the present invention provides a kind of 3D printing method, includes the following steps：
Step A：A kind of 3D printing bio-ink being made of hydrogel and biological cell, the 3D printing biology ink are provided
Water is a kind of gel ink, and the gel ink is added soybean protein isolate solution and stirs under conditions of less than phase transition temperature
It mixes uniformly, calcium sulphate soln is then added and is uniformly mixed into spare 3D printing gel ink material.
Wherein, the gel ink can be the existing 3D printing life being made of hydrogel and biological cell of any type
Object ink, the preferably described gel ink are that its character is suitable for the bio-ink printed using 3D biometric print machines.
Particularly preferably, the gel ink is the material for being exclusively used in the 3D printing method of the present invention, by hydrogel and life
Object cell composition.
3D printing bio-ink example 1
Present example provides a kind of 3D printing bio-ink for the 3D printing method being exclusively used in the present invention, the 3D printing life
Object ink is a kind of gel ink, and the gel ink is made of hydrogel and biological cell, wherein the hydrogel is by as follows
Component is constituted：Cross-linked-hyaluronic acid, poly(N-isopropylacrylamide) (PNIPAAm), poly- (N, N- acrylamide)
(PDEAAm), poly- (2- carboxy-Ns-N-isopropylacrylamide) (PCIPAAm), polymethyl vinyl ether, polyethylene glycol oxidation
Ethylene (PEG-PEO) and deionized water.
In a specific embodiment, the hydrogel may be used following component and constitute：The crosslinking of mass fraction 1% is saturating
Bright matter acid, the poly(N-isopropylacrylamide) of mass fraction 3%, mass fraction 3% poly- (N, N- acrylamide),
Poly- (the 2- carboxy-Ns-N-isopropylacrylamide) of mass fraction 1%, the polymethyl vinyl ether of mass fraction 3%, mass fraction
3% PEG-PEO and the deionized water that surplus is mass fraction 86%.
In another specific embodiment, the hydrogel may be used following component and constitute：The crosslinking of mass fraction 5%
Poly- (N, N- the diethyl acryloyl of hyaluronic acid, the poly(N-isopropylacrylamide) of mass fraction 5%, mass fraction 5%
Amine), poly- (2- carboxy-Ns-N-isopropylacrylamide), the polymethyl vinyl ether of mass fraction 5%, the quality of mass fraction 3%
The PEG-PEO of score 5% and the deionized water that surplus is mass fraction 72%.
In another specific embodiment, the hydrogel may be used following component and constitute：The crosslinking of mass fraction 3%
Poly- (N, N- the diethyl acryloyl of hyaluronic acid, the poly(N-isopropylacrylamide) of mass fraction 4%, mass fraction 4%
Amine)), poly- (2- carboxy-Ns-N-isopropylacrylamide), the polymethyl vinyl ether of mass fraction 4%, the matter of mass fraction 2%
Measure deionized waters of the PEG-PEO of score 4% with surplus for mass fraction 79%.
In there are one specific embodiment, the hydrogel may be used following component and constitute：The friendship of mass fraction 1%
Poly- (N, N- the diethyl acryloyl of connection hyaluronic acid, the poly(N-isopropylacrylamide) of mass fraction 4%, mass fraction 4%
Amine)), poly- (2- carboxy-Ns-N-isopropylacrylamide), the polymethyl vinyl ether of mass fraction 4%, the matter of mass fraction 2%
Measure deionized waters of the PEG-PEO of score 5% with surplus for mass fraction 80%.
Further, the present invention also provides the preparation methods of 3D printing bio-ink described in example 1, including walk as follows
Suddenly：By the cross-linked-hyaluronic acid of mass fraction 1%-5%, the poly(N-isopropylacrylamide) of mass fraction 3%-5%, quality
Poly- (N, the N- acrylamide) of score 3%-5%), the polymethyl vinyl ether of mass fraction 3%-5%, mass fraction
The polyethylene glycol ethylene oxide (PEG-PEO) of 3%-5% is blended with the deionized water of surplus, stirs 30min, stands 1~6
The hydrogel is made in hour；The hydrogel and the biological cell are uniformly mixed into the 3D printing bio-ink.
3D printing bio-ink example 2
Aforementioned exemplary 1 provides one kind and being suitable for the invention 3D printing bio-ink, and the 3D printing bio-ink is
A kind of gel ink, the gel ink are made of hydrogel and biological cell, can be obtained by 3D printing and be supported without holder
Model.Certainly, it will be appreciated by those skilled in the art that using similar gel ink, it can also coordinate with holder ink and beat
Print contains standoff model, of the invention this to be beaten containing two kinds of 3D from existing containing unlike standoff model
Print bio-ink can be through the invention 3D printing method, obtain porous model, can be cell production provide nutrition with
Metabolic pathway is conducive to obtain the thicker institutional framework of bigger by 3D printing.
Specifically, the 3D printing bio-ink of the 3D printing method of the present invention is exclusively used in present example provides another kind,
The 3D printing bio-ink is a kind of superficial degradation type 3D printing bio-ink, by a kind of gel ink and a kind of holder ink
Water is constituted, and the gel ink is made of hydrogel and biological cell, and the holder ink is by the poly- carbonic acid of superficial degradation proximate matter material
Ester lactide copolymer (PTMC-PLLA copolymers) is constituted；The hydrogel of the wherein described gel ink is made of following component：Matter
Measure the PLGA-PEG-PLGA triblock copolymers of score 10%~40%, the cell growth factor of mass fraction 0.001%~1%
Son, the anti-inflammation hemostasia drug of mass fraction 0.05~0.3%, surplus are deionized water；The holder ink is by the gel ink
The makrolon lactide copolymer (PTMC-PLLA copolymers) of the 5%~50% of the gross mass of water is constituted.
3D printing method of the 3D printing bio-ink of the present embodiment especially suitable for the present invention, can pass through 3D printing
One-pass molding obtains the model for having holder to support containing biological cell.That is, this exemplary 3D printing bio-ink can also answer
With the same principle of 3D printing method of the present invention, the wherein preparation of this exemplary gel ink and print procedure and aforementioned exemplary
Gel ink in 1 is identical, and it is solidifying that holder ink uses multi-component material gradient printing type to be formed in the printing of coaxial form of tubes
The inside of celluloid ink water.
More specifically, this exemplary 3D printing bio-ink is utilized when being printed using the 3D methods of the present invention
Two coaxially arranged nozzles are carried out at the same time printing, and specific steps are identical with aforementioned printing step, and different is only to spray
Head is coaxially arranged two, wherein the exemplary holder ink of the nozzle printing sheet being arranged on the inside of coaxitron, such as surface drop
Solution type materials polycarbonate lactide copolymer (PTMC-PLLA copolymers) forms internal stent after printing；It is arranged in coaxial
The gel ink of nozzle printing the present embodiment on the outside of pipe forms the layer structure of tubulose around holder after printing, to profit
With multi-component material gradient printing technique, the double-deck print structure of similar " coaxitron " is constructed.
Above-mentioned 3D printing bio-ink mainly uses PLGA-PEG-PLGA triblock copolymers and PTMC-PLLA copolymerization
Object is as Biodegradable material.The degradation process of biodegradation material macroscopically can behave as superficial degradation and bulk degradation
Two kinds of citation forms, wherein the degradation mode of PLGA-PEG-PLGA triblock copolymers used by the application drops for ontology
The degradation mode of solution, PTMC-PLLA copolymers is superficial degradation.The material surface of bulk degradation mode and internal generation point simultaneously
Solution, dissolving or molecular weight reduce, and it is degradation to cause the strength of materials to decline the characteristics of finally structural collapse, bulk degradation material occur
Speed is unified, degradation time is short.Superficial degradation, which is then material, decomposes on surface, dissolve or molecular weight reduces, and material is successively
Degradation.Superficial degradation mode can guarantee the maintenance of 3D printing biologic material products functional structure in degradation process in human body, such as
Drug release is vector degradation rate control, has special value, PTMC-PLLA copolymers to have surface biological 3D printing product
Degradation characteristic, the PLGA-PEG-PLGA triblock copolymers that can be used with the application coordinate, and can be precisely controlled 3D printing biology
The both macro and micro functional structure of material product, swelling ratio, the degradation rate of controlled material, is conducive to 3D printing biomaterial system
The maintenance of functional structure during product degradation in vivo.
Wherein, PLGA-PEG-PLGA triblock copolymers can be poly- by poly (lactic acid-glycolic acid) (PLGA) and polyethylene glycol (PEG)
It closes and is formed, have application carrying medicine and slow releasing pharmaceutical field, for example, proposing one in Chinese patent application 2015103092137 more
Purposes and preparation method thereof of the kind PLGA-PEG-PLGA triblock copolymers in carrying medicine and slow releasing pharmaceutical, from the prior art
It can be used for carrying the PLGA-PEG-PLGA triblock copolymers of medicine and slow releasing pharmaceutical, the load medicine of the triblock copolymer is molten
Liquid can with temperature increase occur solution-gel phase transition, phase transition temperature be 35~39 DEG C, and the copolymer also have can
Biodegradation character.Load drug solns prepared by the above-mentioned PLGA-PEG-PLGA triblock copolymers proposed in the prior art, work as temperature
When degree is less than phase transition temperature, become solution state；It can then become gel state when temperature reaches phase transition temperature.Therefore drug solns are carried
It is stored in less than being suitable for being injected in solution state under the conditions of human body temperature, after being injected into human body, reaches phase transition temperature and then can
Become gel state, suitable for carrying out degradation release drug in gel state.
Therefore, Thermo-sensitive and degradable characteristic of the present invention using PLGA-PEG-PLGA triblock copolymers, especially carries
A kind of a kind of new application of PLGA-PEG-PLGA triblock copolymers is gone out, being prepared using the triblock copolymer can
Be exclusively used in the present invention 3D printing bio-ink, the 3D printing bio-ink by PLGA-PEG-PLGA triblock copolymers water
Gel and biological cell are constituted.It will be appreciated by those skilled in the art that the preparation in relation to PLGA-PEG-PLGA triblock copolymers
It is referred to the above-mentioned prior art or other existing pertinent literatures with characteristic, can also be adjusted to prepare according to the prior art and be somebody's turn to do
The scheme of triblock copolymer, to obtain suitable phase transition temperature, these are not the scope of protection of the invention, and the present invention is closed
The focus of note is the Thermo-sensitive and degradable characteristic for existing PLGA-PEG-PLGA triblock copolymers, it is proposed that should
Application of the triblock copolymer in 3D printing bio-ink field.
Makrolon lactide copolymer (PTMC-PLLA copolymers) is a kind of degradable high polymer material, due to its tool
Have that good surface erosion degradability, biocompatibility, structure are easily modified, degradation speed is adjustable and the superiority such as workability
Can, it is applied in medicine Disciplinary Frontiers.It is important to note that the makrolon lactide copolymerization of high-carbon acid and esters content
Object superficial degradation is had excellent performance.Due to PTMC-PLLA copolymers have it is unique surface eroding, can avoid material using
In the process, due to largely degrading and causing the drastically decline of mechanical property；PTMC-PLLA copolymers reach 50% in mass loss
When its mechanical strength can still keep 70%~80%.Those skilled in the art can obtain this Shen by the preparation method of the prior art
PTMC-PLLA copolymers please.Same similar, focus of attention of the present invention is for existing PTMC-PLLA copolymers
It can superficial degradation characteristic, it is proposed that using PTMC-PLLA copolymers as internal stent, aforementioned triblock copolymer is as external raw
Long matrix it is hereby achieved that a kind of Thermo-sensitive, having both the 3D printing bio-ink of ontology and superficial degradation characteristic, and is applied to
3D printing field.
For example, those skilled in the art can refer to following existing technical literature, obtains one kind and be suitable for the invention
The preparation method of PTMC-PLLA copolymers.
The existing technical literature is：《The synthesis of Bioabsorbable Polyesters-Copolycarbonate and its structural behaviour are ground
Study carefully》, Fudan University's doctoral thesis, author Yang Jian, 2009, can with specific reference to page 38 page -42 2.1.2.1,
2.1.2.2,2.1.2.4,2.1.3.4 section.
First：Prepare TMC monomers：
The preparation of TMC uses polycondensation-depolymerization process, i.e. 1,3-PD and diethyl carbonate polycondensation under the action of catalyst
De- ethyl alcohol obtains the PTMC of low molecular weight, and then depolymerization cyclization prepares TMC under high temperature action.For example, being connected to fractional distillation column round bottom
76g (1.0mol) l, 3-propanediol, 130g (1.1mol) diethyl carbonate, 50ml dimethylbenzene, 10mg metallic sodiums are added in flask
And 0.1gDBTL, it is heated to flowing back in oil bath, temperature is 140-150 DEG C, collects the second constantly steamed by still at this time
Alcohol (77-8l DEG C of fraction), about 90ml, ester exchange reaction are finished altogether.Remove still, decompression boils off dimethylbenzene and excessive
Diethyl carbonate etc. is volatilizable fraction, for control temperature at 130 DEG C, lg glass puttys are added after 1 hour in reaction, control oil bath temperature 180
DEG C, vacuum distillation obtains white solid crude product, thick yield 85%.With acetone-diethyl ether (1：4) mixed solvent recrystallizes three times,
It is dried under vacuum to constant weight, is stored in phosphorus pentoxide vacuum desiccator.
Secondly：Prepare L- lactide monomers：
The synthesis of L- lactides uses Pfansteihl for raw material, is dehydrated to obtain the polylactic acid of low molecular weight by polycondensation, then
Depolymerization cyclization generates lactide at high temperature, and specific steps are for example：It is slowly risen after Pfansteihl and zinc powder is added in round-bottomed flask
Temperature is used in combination vacuum controller to be gradually decompressed to 10mbar in temperature-rise period complete to dehydration progress to 160-170 DEG C.It replaces and steams
After distillation unit, the L- lactide crude products distilled out at 180-210 DEG C are collected, thick yield is 89%.L- lactides acetic acid second
Ester recrystallizes three times, anhydrous ether washing, then is dried under vacuum to constant weight at normal temperatures, is stored in phosphorus pentoxide vacuum desiccator
In.
Finally：Prepare makrolon lactide copolymer (PTMC-PLLA copolymers)
Using tube sealing ring-opening polymerisation, TMC the and L-LA monomers of different rate of charges are weighed first, the lactic acid of 0.1wt% is added
Zinc catalyst, mixing are added in the glass polymerization pipe of silanization.Heating and melting under nitrogen protection is sufficiently mixed monomer.Ice water
Cooling and solidifying extracts high vacuum in the case where spreading pumping action to 10mbar hereinafter, tube sealing.Glass tube is put into 120 DEG C of oil bath pans, is gathered
Close reaction 72 hours.Gained copolymer is dissolved with dichloromethane, methanol extraction, is finally dried under vacuum to constant weight at 40 DEG C.
Certainly, it will be appreciated by those skilled in the art that above-mentioned preparation method is only a kind of improved based on the prior art
Technical solution, those skilled in the art can also refer to other prior art preparations and obtain the required above-mentioned makrolon of the present invention
Lactide copolymer (PTMC-PLLA copolymers).The emphasis that the present invention is paid close attention to is to utilize makrolon lactide copolymer
The superficial degradation characteristic of (PTMC-PLLA copolymers), to realize the technical purpose of the present invention.
Porcine HGF is a kind of protein molecule that various types of cells can be promoted to be proliferated, and has chemotactic, proliferation and reconstruction
Effect can induce the target cells such as fibroblast, epithelial cell, vascular endothelial cell in inflammatory phase and be moved to damage location, repaiied
Multiple damaged portion；It can promote epithelial cell, vascular endothelial cell, fibroblast proliferation in proliferation period growth factor, promote fine
Tie up cellular rearrangements；The healing of phase acceleration of wound is being rebuild, cicatrization is being reduced, for bone, joint and neurotrosis, cell growth factor
Son can promote skin, subcutaneous tissue, bone tissue reparation and blood vessel, nerve reconstructive, promote new vessels and generate, nerve cell and bone group
Regeneration is knitted, is served in terms of wound healing considerable.
Porcine HGF has basic fibroblast growth factor, epidermal growth factor, insulin-like growth factor, nerve
Trophic factors, transforming growth factor, keratinocyte growth factor, platelet derived growth factor etc..
Basic fibroblast growth factor promotes cell division proliferation and Angiogensis, is to be currently known strongest rush
The Hemapoiesis factor.Fibroblast, epithelial cell, vascular endothelial cell, Skeletal Muscle Cell, cartilage cell, osteoblast are put down
Sliding myocyte, neuron, Deiter's cells, lens epithelial cells, corneal epithelial cell, adrenal cortex medullary epithelium are
The recipient cell of basic fibroblast growth factor.
Epidermal growth factor is synthesized by monocyte and ectoderm cell, stimulate the division of internal multiple types histocyte and
Proliferation, the movement of enhancing cell, merisis and cytoplasm albumen generate, such as skin, cornea, lung, tracheae and liver regeneration.
Insulin-like growth factor is by two close phases of type-1 insulin like growth factor type and insulin-like growth factor 2 type
The family of the small polypeptide composition closed has and promotes muscle protein synthesis, promotes proliferation, the differentiation of sarcoblast, participates in tissue and repair
Multiple and regeneration, to accelerate organization healing.
Neurotrophic factor is present in the trace soluble substance around sensory neuron, is produced by the target cell of neuron
It is raw, specifically promote neure growth and maintenance, promotes tissue damage reparation.
Porcine HGF is introduced into hydrogel by the present invention, and advantage is can be in the bio-ink of subsequent configuration
Cells with nutrient.Holder is printed with 3D printing technique, after stenter to implant human body, cell factor can promote hydrogel
Cell differentiation, increment on holder, accelerate tissue reconstruction.
Sulfa drugs such as sulfapryidine silver, mafenide etc.；Quinolone drugs such as Enoxacin, Norfloxacin, oxygen
Flucloxacillin etc. has antibacterial and anti-inflammation functions.Haemostatic medicament such as adrenobazonum, etamcylate, tranxamic acid, road fourth etc., which has, accelerates blood coagulation
Effect.
In a specific embodiment, the hydrogel may be used following component and constitute：The PLGA- of mass fraction 10%
The sulphur of PEG-PLGA triblock copolymers, the basic fibroblast growth factor of mass fraction 0.01%, mass fraction 0.05%
Amine pyridine silver, surplus are deionized water.
In another specific embodiment, the hydrogel may be used following component and constitute：Mass fraction 15%
PLGA-PEG-PLGA triblock copolymers, the epithelical cell growth factor of mass fraction 1%, the promise fluorine of mass fraction 0.2% are husky
Star, surplus are deionized water.
In another specific embodiment, the hydrogel may be used following component and constitute：Mass fraction 20%
The peace of PLGA-PEG-PLGA triblock copolymers, the keratinocyte growth factor of mass fraction 0.05%, mass fraction 0.25%
Network blood, surplus are deionized water.
In there are one specific embodiment, the hydrogel may be used following component and constitute：Mass fraction 25%
The sulfanilamide (SN) of PLGA-PEG-PLGA triblock copolymers, the insulin-like growth factor of mass fraction 0.5%, mass fraction 0.3%
Myron and etamcylate, surplus are deionized water.
In there are one specific embodiment, the hydrogel may be used following component and constitute：Mass fraction 40%
PLGA-PEG-PLGA triblock copolymers, the neurotrophic growth factor of mass fraction 0.001%, mass fraction 0.15%
Ofloxacin and Lu Ding, surplus are deionized water.
Further, the present invention also provides the preparation method of superficial degradation type 3D printing bio-ink described in example 2, packets
Include following steps：By the PLGA-PEG-PLGA triblock copolymers of mass fraction 10%~40%, mass fraction 0.001%~
1% Porcine HGF, the anti-inflammation hemostasia drug of mass fraction 0.05~0.3% are blended with the deionized water of surplus, stirring
30min stands 1~6 hour and the hydrogel is made；The hydrogel and the biological cell are uniformly mixed into the surface
The gel ink of degradation-type 3D printing bio-ink.
Particularly, in Method of printing of the invention, 3D printing gel ink material controls temperature before carrying out 3D printing
Less than phase transition temperature, gel ink solution is to be in a kind of collosol state of low sticky degree at this time, and approximate colloid passes through addition
Soybean protein isolate solution stirs, and a little calcium sulphate soln is then added, such as add stoste (gel ink) volume 10%
A concentration of 5% soybean protein isolate solution is added the calcium sulphate soln of a concentration of 5-8% of stoste volume 1%-2%, passes through
In sulfuric acid calcium ion and the charge of colloid absorption makes colloid cause cohesion.It is emphasized that this process needs before the printing
It is configured within 15-30 minutes, to prevent coacervation of colloid solidification to be difficult to print.
It needs to be emphasized that the 3D printing gel ink material preferably before the printing 15-30 of step A of the invention
It configures and completes within minute, and printed and completed by printing head in subsequent 15-30 minutes, the bio-ink otherwise configured is held
Easily cohesion solidification blocks nozzle.
In addition, it can be any type vertebrate cells, mammalian cell, people to be suitable for the invention biological cell
Cell or combinations thereof, type depends on produced cell construct, the type of tissue or organ.Such as the cell can
With include be not limited to for shrinkage cell or muscle cell, phoirocyte, bone marrow cell, endothelial cell, Skin Cell,
Epithelial cell, mammary glandular cell, vascular cell, haemocyte, lymphocyte, nerve cell, gastrointestinal tract cell, liver cell, pancreas are thin
Born of the same parents, pneumonocyte, tracheal cell, keratocyte, urogenital cell, nephrocyte, reproduction cell, adipocyte, mesothelial cell,
Cell plastid, entoderm source cell, mesoderm source cell, ectoderm source cell and combinations thereof.In a preferred embodiment, described
Cell is stem cell, including but not limited to embryonic stem cell, adult stem cell, amnion stem cell and inductive pluripotent stem cells
Deng.Wherein the additive amount of biological cell is added according to the difference of different tissues, the speed of growth.
It should be noted that hydrogel specifically preferred according to the invention has temperature-responsive, phase transition temperature range is
20-33 degrees Celsius, when less than phase transition temperature, solution is in low sticky approximation colloidal state.When higher than phase transition temperature, than
Such as 34-37 degrees Celsius, or even to 40 degrees Celsius, hydrogel can be undergone phase transition, gradually cohesion solidification.Be added soy bean proteinous soln and
When calcium sulphate soln, it is necessary to maintain the temperature in transition temperature range and carry out.It is follow-up mixed since hydrogel has temperature-responsive
Bio-ink made of conjunction also has temperature-responsive.It is preferred that phase transition temperature is 25-32 degrees Celsius.More preferably 28-31 takes the photograph
Family name's degree.
The spare 3D printing gel ink material containing gel ink is formd in the step A of the 3D printing method of the present invention
Material, can further be operated later according to step B.
Step B：The temperature for controlling the gel ink sump of 3D biometric print machines is less than phase transition temperature, printing table top control temperature
Degree is 37 degrees Celsius, using the temperature-responsive of gel ink, by 3D printing nozzle by the 3D printing gel ink material
It is printed as model.
In this step, the printer that existing any type is suitable for organism 3D printing may be used in 3D printer, wherein
Aforementioned spare 3D printing gel ink material is placed in gel ink sump, and temperature keeps below phase transition temperature, makes 3D printing
The character of gel ink material is stablized convenient for printing and is not easy to plug nozzle, and the 3D printing nozzle of printer can be arranged and be located at
Printing forms model on printing table top in temperature control box, and in temperature control box.At this point, the temperature in control temperature control box is 37 Celsius
Degree so that model being capable of solidifying and setting.
If it is for the 3D printing bio-ink in aforementioned exemplary 2, used 3D printer may be used existing
It is suitable for 3D printer of the tool there are two coaxially arranged nozzle of organism 3D printing, there are one gel inks for 3D printer tool
Sump and a holder ink storehouse, printing step and the principle of abovementioned steps B are essentially identical, i.e.,：Control 3D biometric print machines
The temperature of gel ink sump is less than phase transition temperature, and printing table top temperature is 37 degrees Celsius, and using tool, there are two coaxially arranged
The 3D printer of nozzle, holder ink described in the inner circumferential side nozzle printing by the 3D printer forms holder, by described
The 3D printing gel ink material is printed as model by the peripheral side nozzle of 3D printer around the holder.Certainly, it prints
In the process, gel ink is the nozzle printing being arranged on the outside of coaxitron by 3D printer, and holder ink is to pass through 3D
The nozzle printing of printer being arranged on the inside of coaxitron.
It should be noted that be added to soybean protein isolate in abovementioned steps molten for gel ink specifically preferred according to the invention
Liquid and calcium sulphate soln have carried out pre-agglomeration, agglomerate solidification after being printed as model.The variation and the regulation and control of time of temperature make biology
Ink has two kinds of coacervation process, and when being printed on temperature control box printing table top by printing head, the droplet printed every time is very
It is easy to form fixed structure, thus the above-mentioned particularly preferred gel ink of the present invention may not need covering after one layer of printing
One layer of timbering material is supported, and provides a kind of no holder 3D biometric print technologies.Above-mentioned 3D namely through the invention
The step A and step B of Method of printing, can directly obtain the 3D printing model supported without holder on the basis of hydrogel, can
To save the time-write interval of holder, without print head is needed to change, can save time-write interval and cost.
Certainly, be not precluded can be by existing 3D printer successively printing alternate bio-ink and holder material by the present invention
Material forms the technical solution of model, that is, in order to form complicated model structure, in a preferred embodiment, this step is into one
Step include in temperature control box by 3D printing nozzle by the bio-ink and timbering material successively printing alternate at the model,
To provide support to bio-ink by timbering material.Wherein, it is existing suitable can be selected from any type for the timbering material
For the timbering material of 3D printing, including be not limited to fibrin, alginates, agarose, chitosan and combinations thereof.
Alternatively, as described in aforementioned exemplary 2, gel ink of the invention can also pass through the 3D printing of coaxially arranged nozzle
It is holder that machine, which is obtained with the inner circumferential side of holder ink cooperation, the structural model that peripheral side is gel growth matrix.
Wherein, the preparation of the gel ink of peripheral side and print conditions are identical as the print procedure of no rack structure model,
The melting temperature of the holder ink of inner circumferential side, for example, makrolon lactide copolymer (PTMC-PLLA copolymers) melting temperature
Degree is 100 degrees Celsius or more, and therefore, holder ink storehouse needs to keep the condition of high temperature when printing, by low after printing
Temperature is formed by curing holder.The gel ink and holder ink that the example 2 of the present invention uses have the two kinds of phase transition completely contradicted
Process, gel ink needs are printed with liquid under low-temperature condition, are cured by 37 degrees Celsius of table top temperature；And holder
Ink needs form printable flow-like under 100 degrees Celsius or more of the condition of high temperature, while in 37 degrees Celsius of table top temperature
Degree is cured.And coaxitron gradient printing type of the present invention, it can be when two nozzles work, using interior
The high temperature holder ink of side accelerates the low temperature gel ink solidification in outside, while the low temperature gel ink in outside being utilized to accelerate inside
High temperature holder ink solidification.Thus, the 3D printing ink of example 2 of the invention can substantially reduce the control of table top temperature
Precision, curing rate faster, printing effect also higher.
Further, the present invention can also be right on the basis of above-mentioned steps A and step B by following additional step
Model carries out subsequent processing, to generate a kind of porous model structure.
Step C：The model is placed in temperature control box (if model is directly printed upon in temperature control box, can be omitted by
Model is placed in the step of temperature control box), the temperature in the temperature control box is slowly reduced to 0 degree hereinafter, making the moisture in the model
It freezes completely.The effect of this step is so that the water in model is build-up ice by cooling, expands 10% from the water volume of maximal density,
By big hole of the formation containing ice of support inside the jellium model of cohesion.Should it is especially mentioned that, the process of cooling needs very slow
Slowly, it is ensured that by liquid slow coagulation, water in model structure is evenly dispersed to congeal into ice, and keeps model as possible for water in model
Internal structure, so that model overall structure is destroyed, ice-out is difficult to keep tissue morphology later for the too fast meeting that otherwise cools down.It is preferred that
In this step, the temperature in the temperature control box is slowly reduced to subzero 15 to minus 20 degrees with the speed of 4-5 degree per hour.
It should be strongly noted that when temperature is reduced to subzero, if not doing protection processing, contain in biological cell
Moisture can also freeze, the ice crystal of formation can the basic structure of damaging cells lead to biological cell loss of activity.Therefore, it to carry out
The refrigerating process of step C needs in aforementioned step A, before biological cell is prepared into 3D printing bio-ink, to life
Object cell carries out protection processing：Biological cell and a concentration of 10% glycerine and a concentration of 10% starch solution are uniformly mixed
It closes and stands 2-3 hours.The effect of this step is to be wrapped up biological cell using the colloid of starch solution, is reduced using glycerine
The freezing point of moisture in biological cell, under the conditions of the slow freezing of step C, intracellular water appears, and reduces ice crystal and is formed,
To avoid cellular damage, ensure biological cell survival.In a specific embodiment, the glycerine of concentration 10% and a concentration of
The additive amount of 10% starch solution can be respectively 1-1.5 times and 0.5-0.8 times of biological cell quality.
In addition, the temperature-fall period of the present invention will not cause model to cave in.There are two types of agglomerated bio-ink tool of the present invention
Journey, one is the cohesion of the temperature-responsive of 3D printing gel ink material, one is be added after soybean protein and calcium sulfate when
Between control agglomerate.In temperature-fall period, moisture can be very good to seek connections in soybean protein and sulphur 3D biomaterials before freezing
Sour calcium agglomerates on the skeleton to be formed, and is unlikely to cave in.
Step D：The temperature in the temperature control box is quickly increased to 37 degrees Celsius, keeps the hydrogel in the model completely solid
Change, to obtain the porous structural model containing biological cell.In this step, model slow cooling to subzero 15 to subzero 20
After degree, in the case where model volume is usually little, the water in model all condenses, at this time can be by being rapidly heated
To 37 degree of organism active temperature, the hydrogel in model is made to cure again, while the ice-out in model, in gel solidification
During moisture deviate from, the hole containing ice can remain in cured gel structure, so as to form biological cell is contained
Porous structure.
Particularly, the temperature-rise period in this step is different from temperature-fall period above-mentioned, need as early as possible lift scheme temperature make
It obtains structure as early as possible to cure, solidification is avoided slowly hole to be caused to disappear.It in a preferred embodiment, can be to institute in this step
The dry-heat air for stating 37 degree of conveying in temperature control box, makes the ice rapid translating in the model be escaped from the model at gas, from
And form porous structure on the model.Dry-heat air can promote the temperature in temperature control box to 37 degree quickly, dry simultaneously
The moisture in temperature control box has been discharged in hot wind, reduces water vapor partial pressure, has further speeded up the speed that moisture is converted into gas, thus
The present embodiment can achieve the effect that model is made to be formed by curing porous structure as early as possible.
After the porous structure containing biological cell obtained on model, model can be placed in nutritive solution and be carried out
Culture obtains the channel of nutrition and metabolism since porous structure provides for biological cell, thus the cell inside model is the same as outer
Portion's cell equally can obtain enough nutrient growth, and this 3D printing method acquisition bigger that can be through the invention is thicker
Institutional framework.
After being closed in order to avoid institutional framework growth, the easy missing nutrient of internal cell, metabolism are unsmooth and dead, another
In one preferred embodiment, after obtaining the porous structural model containing biological cell, injected in the porous structure
Vascular endothelial cell makes vascular endothelial cell be attached on structure pore framework, model is placed in nutritive solution carries out later
Culture, goes out blood vessel by vascular endothelial cell growth, and original cell growth is at required tissue in model, the blood vessel that grows
Further nutrient and metabolic pathway are provided for required tissue.Further, in order to avoid the hole inside porous structure does not connect
It is logical, cause the blood vessel of growth to be difficult to the defect penetrated through, in another preferred embodiment, biological cell can be contained obtaining
After porous structural model, the model is uniformly got to the parallel via holes of perforation by laser, is used for the hole in model
Hole connection is got up.The through-hole diameter that laser boring obtains is limited, be appropriate only for hole connection convenient for vascular endothelial cell and
Nutrient solution enters inner void, is operated according to aforesaid way thereafter, and vascular endothelial cell is injected in porous structure, makes intravascular
Chrotoplast is attached on structure pore framework, model is placed in nutritive solution cultivates later, pass through vascular endothelial cell
Grow blood vessel.With the growth of blood vessel and tissue, the through-hole for these perforations that laser boring obtains can be stretched, when
So, adequate thickness and the tissue of volume are obtained, or the main porous structure mould for needing to obtain in dependence previous embodiment
Type.
It will be appreciated by those skilled in the art that although the present invention is described in the way of multiple embodiments,
It is that not each embodiment only contains an independent technical solution.So narration is used for the purpose of for the sake of understanding in specification,
The skilled in the art should refer to the specification as a whole is understood, and by technical solution involved in each embodiment
Regard as and can be combined with each other into the mode of different embodiments to understand protection scope of the present invention.
The foregoing is merely the schematical specific implementation modes of the present invention, are not limited to the scope of the present invention.It is any
Those skilled in the art, do not depart from the design of the present invention and under the premise of principle made by equivalent variations, modification and combination,
The scope of protection of the invention should all be belonged to.
Claims (4)
1. a kind of 3D printing method of superficial degradation type 3D printing bio-ink, wherein the superficial degradation type 3D printing biology
Ink is made of a kind of gel ink and a kind of holder ink, and the gel ink is made of hydrogel and biological cell, described
Hydrogel is made of following component：PLGA-PEG-PLGA triblock copolymers, the mass fraction of mass fraction 10%~40%
0.001%~1% Porcine HGF, the anti-inflammation hemostasia drug of mass fraction 0.05~0.3%, surplus are deionized water；It is described
Holder ink is made of 5%~50% makrolon lactide copolymer of the gross mass of the gel ink；
The 3D printing method includes the following steps：
Step A：The gel ink is added soybean protein isolate solution and stirs evenly under conditions of less than phase transition temperature,
Then calcium sulphate soln is added and is uniformly mixed into spare 3D printing gel ink material；
Step B：The temperature for controlling the gel ink sump of 3D biometric print machines is less than phase transition temperature, and printing table top temperature is taken the photograph for 37
Family name's degree passes through the inner circumferential side nozzle printing institute of the 3D printer using tool there are two the 3D printer of coaxially arranged nozzle
It states holder ink and forms holder, the 3D printing gel ink material is surrounded by institute by the peripheral side nozzle of the 3D printer
It states holder and is printed as model；
Step C：The model is placed in temperature control box, slowly reduces the temperature in the temperature control box to 0 degree hereinafter, making the mould
Moisture in type freezes completely；
Step D：The temperature in the temperature control box is quickly increased to 37 degrees Celsius, the hydrogel in the model is made to be fully cured,
Obtain the porous structural model containing biological cell；
After obtaining the porous structural model containing biological cell, the model is uniformly got into perforation by laser
Parallel via holes, then inject vascular endothelial cell in the porous structure, the vascular endothelial cell made to be attached to structure
On pore framework, the model is placed in nutritive solution cultivates later, passes through the vascular endothelial cell growth bleeding
Pipe, at required tissue, the blood vessel that grows further provides for required tissue foster original cell growth in the model
Point and metabolic pathway.
2. 3D printing method as described in claim 1, which is characterized in that in the step A, preparing the 3D printing biology
Before ink, protection processing is carried out to the biological cell：By the biological cell and a concentration of 10% glycerine and a concentration of
10% starch solution uniformly mixes standing 2-3 hours.
3. 3D printing method as described in claim 1, which is characterized in that in the step C, with the speed of 4-5 degree per hour
Temperature in the temperature control box is slowly reduced to subzero 15 to minus 20 degrees.
4. 3D printing method as described in claim 1, which is characterized in that in the step D, 37 are conveyed into the temperature control box
The dry-heat air of degree makes the ice rapid translating in the model be escaped from the model at gas, is formed on the model more
Pore structure.
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| CN201710790405.3A Active CN107523136B (en) | 2016-12-09 | 2017-09-05 | Temperature-responsive degradable 3D printing biological ink and 3D printing method |
| CN201710790467.4A Active CN107400412B (en) | 2016-12-09 | 2017-09-05 | A kind of polyestercarbonate acid anhydrides 3D printing bio-ink and 3D printing method |
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| CN108126244A (en) * | 2018-02-27 | 2018-06-08 | 深圳大学 | A kind of tissue engineering bracket and preparation method thereof and it is a kind of can direct biomineralization 3D printing ink |
| CN109395151A (en) * | 2018-10-25 | 2019-03-01 | 浙江大学 | A kind of preparation method and application of 3D printing hemostatic material |
| CN109731130B (en) * | 2018-11-14 | 2021-09-24 | 华中科技大学同济医学院附属协和医院 | Method for preparing hydrogel wound dressing by low-temperature biological 3D printing technology |
| KR20220060501A (en) * | 2019-03-29 | 2022-05-11 | 티디비티 아이피 인코포레이티드 | Tissue and organ substitutes and methods of making them |
| CN112917891B (en) * | 2019-12-06 | 2022-08-02 | 苏州永沁泉智能设备有限公司 | Biological 3D printing method for massive tissues |
| CN112917892B (en) * | 2019-12-06 | 2022-08-02 | 苏州永沁泉智能设备有限公司 | Biological 3D printing method based on nutrition flow channel |
| CN112917899B (en) * | 2019-12-06 | 2023-01-06 | 苏州永沁泉智能设备有限公司 | Biological 3D printing method based on porous structure |
| CN110982335A (en) * | 2019-12-30 | 2020-04-10 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of self-curing hydroxyapatite 3D printing ink |
| CN111388750B (en) * | 2020-04-30 | 2022-09-13 | 深圳先进技术研究院 | Biological ink, small-caliber tubular structure support and preparation method and application thereof |
| GB202103785D0 (en) * | 2021-03-18 | 2021-05-05 | UCB Biopharma SRL | Formulations |
| CN113180879B (en) * | 2021-05-24 | 2023-12-22 | 上海大学 | Biological tissue preparation system and method |
| CN114134731A (en) * | 2021-09-06 | 2022-03-04 | 苏州大学 | Direct-injection acid dye digital printing ink and application thereof |
| CN114059362A (en) * | 2021-11-17 | 2022-02-18 | 江苏博来特新材料有限公司 | Thermosensitive gelation direct injection type textile ink and application thereof |
| CN114259604B (en) * | 2021-12-17 | 2022-12-27 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of 3D printing ordered vascularization promoting drug-loaded bone repair scaffold, product and application thereof |
| CN114395930A (en) * | 2021-12-31 | 2022-04-26 | 山东中康国创先进印染技术研究院有限公司 | Direct injection type reactive dye ink used for printed fabric and free of front and back treatment and application thereof |
| EP4349778A1 (en) * | 2022-10-03 | 2024-04-10 | Universitat Internacional De Catalunya, Fundació Privada | Ink for 3d printing |
| CN115957374A (en) * | 2022-12-16 | 2023-04-14 | 吉林大学 | Metal artificial bone implant with core-shell structure and preparation method thereof |
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| JP6521328B2 (en) * | 2014-02-28 | 2019-05-29 | 国立大学法人大阪大学 | Method for producing a three-dimensional structure of cells |
| CN103977453A (en) * | 2014-05-28 | 2014-08-13 | 段升华 | 3D (three-dimensional) bioprinting hydrogel material and application thereof |
| CN104511058A (en) * | 2014-12-29 | 2015-04-15 | 复旦大学 | Fully bio-absorbable polymer stent and production method thereof |
| CN105017495B (en) * | 2015-06-29 | 2017-11-10 | 西北师范大学 | A kind of preparation of soybean protein base dual-sensitivity high molecule micropore gel and the application as pharmaceutical carrier |
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Address after: 310018 room 2, building 452, No. 6, main street, Hangzhou economic and Technological Development Zone, Zhejiang, Patentee after: Hangzhou Mingshan Biotechnology Co., Ltd Address before: 310018 room 2, building 452, No. 6, main street, Hangzhou economic and Technological Development Zone, Zhejiang, Patentee before: Hangzhou Medzone Bio-Technology Co., Ltd. |
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