CN107400412A - A kind of polyestercarbonate acid anhydrides 3D printing bio-ink and 3D printing method - Google Patents
A kind of polyestercarbonate acid anhydrides 3D printing bio-ink and 3D printing method Download PDFInfo
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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 polyestercarbonate acid anhydrides 3D printing bio-ink and 3D printing method, the 3D printing bio-ink is made up of a kind of gel ink and a kind of support ink, the gel ink is made up of hydrogel and biological cell, the hydrogel includes the PLGA PEG PLGA triblock copolymers with temperature-responsive, and the support ink is the polyestercarbonate acid anhydride copolymer (P (LLA TMC SA) copolymer) 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, multi-component material gradient printing type is used to obtain the model for possessing controlled surface degraded support support containing biological cell with the printing of coaxial form of tubes, and the nutrition and metabolism passage of loose structure can be obtained, is advantageous to obtain bigger thicker institutional framework by 3D printing.
Description
Technical field
The present invention relates to biological technical field, especially 3D biometric prints technical field, more particularly to a kind of superficial degradation
Type polyestercarbonate acid anhydrides 3D printing bio-ink and 3D printing method.
Background technology
3D printing can be rapidly and efficiently the product for producing personalization, 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 application to 3D biometric print technologies be divided into different levels progress
Introduce, 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
In biodegradable support hold and last, then cultivated in bioreactor so that cell relies on support to be formed
Spatial growth generation needed for organize.For example, the A of CN 106085949 disclose a kind of reconstruction urethra based on 3D printing shaping
The method of prosthese；Epithelioid cell, class smooth muscle cell are divided into by cell inducible factors, is then converted into cell drop,
The cell drop has cell and nutrient solution hydrogel to mix, and the bio-ink for being available for medical 3D printer to use is made；Choosing
Select the support ink of hydrogel that collagen mixes 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, lifted with electrical-controlled lifting platform courses shower nozzle, 3D printing
Two shower 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, cell drop is then converted into, with reference to 3D printing, rebuilds urethra in vitro, then will rebuild again
Urethra be transplanted to patient with it go.That is, the 3D biometric prints material used in above-mentioned existing 3D biometric prints technology is two
Kind：One kind is to wrap celliferous bio-ink, a kind of timbering material for being used as support.
Prior art is basically to find suitable bio-ink and timbering material, example on the research direction of 3D printing
Such as：The A of CN 104399119 disclose a kind of method that strong mechanical performance cartilage is prepared based on 3D biometric prints, and it passes through silk
Fiber and gelatin solution prepare the bio-ink containing cartilage stem cell, and forming support 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 the ring-opening polymerisation of trihydroxy initiator2-(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 light triggers 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 is molded after completing by UV light irradiations.For another example：CN
A kind of bio-ink for 3D printing is disclosed in 105238132 A, its constituent is included with the water-soluble of crosslinking function
Property synthetic polymer, there is the water-soluble natural macromolecule of 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 UV light irradiations
Type.The A of CN 105885436 disclose a kind of bio-ink material for 3D printing and its preparation method and application, and this is existing
Technology provides timbering material in fact, and the timbering material is used as support ink by the use of large biological molecule, precrosslinker, coagulant
Material, shape after printing by washing, crosslinking agent, acquisition is antigenic, rejection is small and has biodegradability
Soft tissue support.
Described above is the background of the 3D biometric print technologies comprising support, disclosed in the A of CN 103249567
Be used for manufacture in the devices, systems, and methods of tissue, for the various materials of 3D biometric prints component carried out it is detailed
Enumerate, wherein specifically mentioned prior art also whether there is the trial of support 3D biometric print technologies, but exist very without support technology
More limitations, such as be difficult to obtain complicated geometry, be difficult to be formed blood vessel network of nutrient etc. needed for offer tissue production.Cause
This, what the prior art provided is the 3D biometric print solutions comprising support.
In fact a very thin layer tissue can only be printed just because of above-mentioned reason, existing 3D biometric prints technology, because
For compared with, without nutrition channels such as blood vessels, internal cell is difficult to acquisition nutriment inside thick tissue, metabolite can not discharge,
Thus thicker 3D printing tissue is difficult to Sustainable Growth.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of superficial degradation type polyestercarbonate acid anhydrides 3D printing biology ink
Water and 3D printing method, so that the problem of being formerly mentioned is reduced or avoided.
Specifically, the invention provides a kind of superficial degradation type polyestercarbonate acid anhydrides 3D printing bio-ink and 3D to beat
Impression method, the demand maintained for structure-controllable in degradation process in 3D biometric print stake bodies, preparing has superficial degradation special
Property printable bioabsorbable polymer material, can print obtain containing biological cell possess controlled degradation support support mould
Type, and porous model can also be further obtained, it can be produced for cell and nutrition and metabolism passage is provided, be advantageous to pass through
3D printing obtains bigger thicker institutional framework.
In order to solve the above technical problems, the present invention proposes a kind of superficial degradation type polyestercarbonate acid anhydrides 3D printing biology
Ink, it is made up of a kind of gel ink and a kind of support ink, the gel ink is made up of hydrogel and biological cell, its
In, the hydrogel is made up of following component：The PLGA-PEG-PLGA triblock copolymers of mass fraction 10%~40%, matter
Amount the Porcine HGF of fraction 0.001%~1%, the anti-inflammation hemostasia medicine of mass fraction 0.05~0.3%, surplus for go from
Sub- water；The support ink is made up of 5%~50% polyestercarbonate acid anhydride copolymer of the gross mass of the gel ink.
Further, present invention also offers a kind of above-mentioned superficial degradation type polyestercarbonate acid anhydrides 3D printing bio-ink
Preparation method, comprise the following steps：By the PLGA-PEG-PLGA triblock copolymers of mass fraction 10%~40%, quality
The Porcine HGF of fraction 0.001%~1%, the anti-inflammation hemostasia medicine of mass fraction 0.05~0.3% and surplus go from
Sub- water blending, stirs 30min, stands 1~6 hour and the hydrogel is made；The hydrogel and the biological cell is uniform
It is mixed into the gel ink of the 3D printing bio-ink.
In addition, present invention also offers one kind to use above-mentioned superficial degradation type polyestercarbonate acid anhydrides 3D printing bio-ink
3D printing method, comprise the following steps：
Step A：Using above-mentioned superficial degradation type polyestercarbonate acid anhydrides 3D printing bio-ink, the gel ink is low
Under the temperature conditionss of phase in version, add soybean protein isolate solution and stir, then add calcium sulphate soln and uniformly mix
Into standby 3D printing gel ink material；
Step B：The temperature of the gel ink sump of 3D biometric print machines is controlled to be less than phase transition temperature, printing table top temperature is
37 degrees Celsius, using the 3D printer with two coaxially arranged shower nozzles, beaten by the inner circumferential side shower nozzle of the 3D printer
Print the support ink and form support, enclosed the 3D printing gel ink material by the outer circumferential side shower nozzle of the 3D printer
Model is printed as around the support.
Preferably, in the step A, the biological cell is before the 3D printing bio-ink is prepared, to the life
Thing cell carries out protection processing：The starch solution that the glycerine and concentration for being 10% with concentration by the biological cell are 10% 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, the temperature in the temperature control box is slowly reduced to less than 0 degree, makes institute
The moisture stated in model freezes completely；
Step D：The temperature in the temperature control box is quickly raised to 37 degrees Celsius, makes the hydrogel in the model completely solid
Change, so as to obtain the porous structural model containing biological cell.
Preferably, in the step C, temperature in the temperature control box is slowly reduced to zero with the speed of 4-5 degree per hour
15 are descended 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 escapes into gas from the model, so as to form loose structure on the model.
Preferably, methods described further comprises, is obtaining the porous structural model containing biological cell
Afterwards, vascular endothelial cell is injected in the loose structure, the vascular endothelial cell is attached on structure pore framework,
The model is placed in nutritive solution afterwards and cultivated, blood vessel, the model are gone out by the vascular endothelial cell growth
In original cell growth into required tissue, the blood vessel grown further provides nutrient for required tissue and metabolism is logical
Road.
Preferably, methods described further comprises：Obtaining the porous structural model containing biological cell
Afterwards, the model is uniformly got to the parallel via holes of insertion by laser, for the hole connection in model to be got up, then
Vascular endothelial cell is injected in the loose structure, the vascular endothelial cell is attached on structure pore framework, afterwards
The model is placed in nutritive solution and cultivated, blood vessel, the model Central Plains are gone out by the vascular endothelial cell growth
For some cell growths into required tissue, the blood vessel grown further provides nutrient and metabolic pathway for required tissue.
The superficial degradation type polyestercarbonate acid anhydrides 3D printing bio-ink of the present invention is by gel ink and support ink structure
Into wherein gel ink is made up of hydrogel and biological cell, and the hydrogel includes the PLGA-PEG- with temperature-responsive
PLGA triblock copolymers, the support ink are the polyestercarbonate acid anhydride copolymer with superficial degradation characteristic.The present invention
Using above-mentioned 3D printing bio-ink Thermo-sensitive and can superficial degradation characteristic, for being degraded in 3D biometric print stake bodies
The demand that structure-controllable maintains in journey, the printable bioabsorbable polymer material with superficial degradation characteristic is prepared, can once be beaten
It is printed as type and obtains the model for possessing controlled degradation support and supporting containing biological cell, and the nutrition of loose structure can be obtained
And metabolic pathway, be advantageous to obtain bigger thicker institutional framework by 3D printing.
Embodiment
In order to which technical characteristic, purpose and the effect of the present invention is more clearly understood, now pass through the specific of the present invention
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 support
Material, the defects of being difficult to overcome the tissue of printing acquisition to lack nutrition and metabolism passage.Therefore, the invention provides a kind of new
3D printing method, its can by 3D printing obtain containing biological cell without support support or one-shot forming have support support
Model, and can also further obtain porous model, can be produced for cell and nutrition and metabolism passage be provided, be advantageous to
Bigger thicker institutional framework is obtained by 3D printing.
Specifically, the present invention provides a kind of 3D printing method, comprises the following steps：
Step A：A kind of 3D printing bio-ink being made up of hydrogel and biological cell, the 3D printing biology ink are provided
Water is a kind of gel ink, and the gel ink adds soybean protein isolate solution and stirred under conditions of less than phase transition temperature
Mix uniformly, then add calcium sulphate soln and be uniformly mixed into standby 3D printing gel ink material.
Wherein, the gel ink can be any existing 3D printing life being made up of hydrogel and biological cell
Thing ink, preferably described gel ink are the bio-ink that its character is suitable for being 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, and it is by hydrogel and life
Thing cellularity.
3D printing bio-ink example 1
Present example provides a kind of 3D printing bio-ink for the 3D printing method for being exclusively used in the present invention, the 3D printing life
Thing ink is a kind of gel ink, and the gel ink is made up of hydrogel and biological cell, wherein the hydrogel is by as follows
Component is formed：Cross-linked-hyaluronic acid, NIPA (PNIPAAm), poly- (N, N- acrylamide)
(PDEAAm), poly- (2- carboxy-Ns-N-isopropylacrylamide) (PCIPAAm), polymethyl vinyl ether, polyethylene glycol oxidation
Ethene (PEG-PEO) and deionized water.
In a specific embodiment, the hydrogel can use following component to form：The crosslinking of mass fraction 1% is saturating
Bright matter acid, the NIPA 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 can use following component to form：The crosslinking of mass fraction 5%
Hyaluronic acid, the NIPA of mass fraction 5%, poly- (N, N- the diethyl acryloyl of 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 fraction 5% and the deionized water that surplus is mass fraction 72%.
In another specific embodiment, the hydrogel can use following component to form：The crosslinking of mass fraction 3%
Hyaluronic acid, the NIPA of mass fraction 4%, poly- (N, N- the diethyl acryloyl of mass fraction 4%
Amine), poly- (2- carboxy-Ns-N-isopropylacrylamide), the polymethyl vinyl ether of mass fraction 4%, the quality of mass fraction 2%
The PEG-PEO of fraction 4% and the deionized water that surplus is mass fraction 79%.
In an also specific embodiment, the hydrogel can use following component to form：The friendship of mass fraction 1%
Join hyaluronic acid, the NIPA of mass fraction 4%, poly- (N, N- the diethyl acryloyl of mass fraction 4%
Amine), poly- (2- carboxy-Ns-N-isopropylacrylamide), the polymethyl vinyl ether of mass fraction 4%, the quality of mass fraction 2%
The PEG-PEO of fraction 5% and the deionized water that surplus is mass fraction 80%.
Further, present invention also offers the preparation method of 3D printing bio-ink described in example 1, including following step
Suddenly：By mass fraction 1%-5% cross-linked-hyaluronic acid, mass fraction 3%-5% NIPA, quality
Fraction 3%-5% poly- (N, N- acrylamide), mass fraction 3%-5% polymethyl vinyl ether, mass fraction
3%-5% polyethylene glycol ethylene oxide (PEG-PEO) 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 a kind of 3D printing bio-ink for being applied to the present invention, and the 3D printing bio-ink is
A kind of gel ink, the gel ink are made up of hydrogel and biological cell, can be obtained by 3D printing and be supported without support
Model.Certainly, it will be appreciated by those skilled in the art that using similar gel ink, it can also coordinate with support ink and beat
Print contain standoff model, contains from existing unlike standoff model, of the invention this to be beaten containing two kinds of 3D
Print bio-ink can by the present invention 3D printing method, obtain porous model, can be cell production provide nutrition with
Metabolic pathway, be advantageous to obtain bigger thicker institutional framework 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 polyestercarbonate acid anhydrides 3D printing bio-ink, and it is by a kind of gel ink
Water and a kind of support ink are formed, and the gel ink is made up of hydrogel and biological cell, and the support ink is dropped by surface
Section bar material polyestercarbonate acid anhydride copolymer (P (LLA-TMC-SA) copolymer) is solved to form；The water-setting of wherein described gel ink
Glue is made up 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 medicine of mass fraction 0.05~0.3%, surplus are deionized water；
The support ink by the gel ink gross mass 5%~50% polyestercarbonate acid anhydride copolymer (P (LLA-
TMC-SA) copolymer) form.
3D printing method of the 3D printing bio-ink of the present embodiment especially suitable for the present invention, it can pass through 3D printing
One-shot forming obtains the model for having support to support containing biological cell.That is, the 3D printing bio-ink of this example can also answer
With the same principle of the 3D printing method of the present invention, the wherein preparation of the gel ink of this example and print procedure and aforementioned exemplary
Gel ink in 1 is identical, and support ink uses multi-component material gradient printing type to print to be formed solidifying with coaxial form of tubes
The inside of celluloid ink water.
More specifically, the 3D printing bio-ink of this example utilizes when the 3D methods printing using the present invention
Two coaxially arranged shower nozzles are printed simultaneously, and its specific steps and foregoing printing step identical, different is only to spray
Head is coaxially arranged two, wherein the support ink of nozzle printing this example being arranged on the inside of coaxitron, such as surface drop
Section bar material polyestercarbonate acid anhydride copolymer (P (LLA-TMC-SA) copolymer) is solved, internal stent is formed after printing；It is arranged in
The gel ink of nozzle printing the present embodiment on the outside of coaxitron, the layer structure of tubulose is formed after printing around support, from
And multi-component material gradient printing technique is utilized, construct the double-deck print structure of similar " coaxitron ".
Above-mentioned 3D printing bio-ink mainly employs PLGA-PEG-PLGA triblock copolymers and P (LLA-TMC-SA)
Copolymer is as Biodegradable material.The degradation process of biodegradation material macroscopically can behave as superficial degradation and body
Two kinds of citation forms of degraded, wherein, the degraded mode of PLGA-PEG-PLGA triblock copolymers is body used by the application
Degraded, the degraded mode of P (LLA-TMC-SA) copolymer is superficial degradation.The material surface of bulk degradation mode and inside are simultaneously
Decompose, dissolve or molecular weight reduce, cause the strength of materials decline finally there is the characteristics of structural collapse, bulk degradation material
It is that degradation speed is unified, degradation time is short.Superficial degradation be then material decomposed on surface, dissolve or molecular weight reduce, material
Material is successively degraded.Superficial degradation mode can ensure the dimension of 3D printing biologic material products functional structure in degradation process in human body
Hold, if insoluble drug release is vector degradation speed control, there is special value, P (LLA-TMC-SA) copolymerization to biological 3D printing product
Thing has superficial degradation characteristic, and the PLGA-PEG-PLGA triblock copolymers that can be used with the application coordinate, and can be precisely controlled
The both macro and micro functional structure of 3D printing biologic material products, swelling ratio, the degradation rate of controlled material, is advantageous to 3D printing
The maintenance of functional structure during biologic material products degradation in vivo.
Wherein, PLGA-PEG-PLGA triblock copolymers can be poly- by poly (lactic acid-glycolic acid) (PLGA) and polyethylene glycol (PEG)
Close and formed, have application carrying medicine and slow releasing pharmaceutical field, for example, proposing one in Chinese patent application 2015103092137 more
The purposes of kind PLGA-PEG-PLGA triblock copolymers in medicine and slow releasing pharmaceutical is carried and preparation method thereof, from the prior art
It can obtain molten available for the PLGA-PEG-PLGA triblock copolymers for carrying medicine and slow releasing pharmaceutical, the load medicine of the triblock copolymer
Liquid can with temperature raise occur solution-gel phase in version, 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, it is changed into solution state；It can then be changed into gel state when temperature reaches phase transition temperature.Therefore drug solns are carried
It is stored in and is less than under the conditions of human body temperature suitable for being injected in solution state, after being expelled to human body, reaches phase transition temperature and then can
It is changed into gel state, suitable for carrying out degraded release medicine in gel state.
Therefore, Thermo-sensitive and degradable characteristic of the present invention using PLGA-PEG-PLGA triblock copolymers, is especially carried
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 formed.It will be appreciated by those skilled in the art that the preparation about PLGA-PEG-PLGA triblock copolymers
Above-mentioned prior art or other existing pertinent literatures are referred to characteristic, can also be adjusted to prepare according to 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.
Polyestercarbonate acid anhydride copolymer (P (LLA-TMC-SA) copolymer) is a kind of degradable high polymer material, due to
It has that good surface erosion degradability, biocompatibility, structure are easily modified, degradation speed is adjustable and workability etc. is excellent
Performance, it is applied in medical science Disciplinary Frontiers.Because P (LLA-TMC-SA) copolymer has the surface eroding of uniqueness, its
Material can be avoided in use, cause the drastically decline of mechanical property due to a large amount of degradeds；P (LLA-TMC-SA) is copolymerized
Thing its mechanical strength when mass loss reaches 60% can still keep 70%~80%.Those skilled in the art can be by existing skill
The preparation method of art obtains P (LLA-TMC-SA) copolymer of the application.Same similar, focus of attention of the present invention is to be directed to
Existing P (LLA-TMC-SA) copolymer can superficial degradation characteristic, it is proposed that using P (LLA-TMC-SA) copolymers as internal
Support, foregoing triblock copolymer is as external growth matrix, it is hereby achieved that a kind of Thermo-sensitive, having body and surface drop concurrently
The 3D printing bio-ink of characteristic is solved, and is applied to 3D printing field.
For example, those skilled in the art may be referred to any prior art literature, obtain a kind of suitable for the present invention
P (LLA-TMC-SA) copolymer preparation method.Or the present invention can also prepare the P of the present invention with the following method
(LLA-TMC-SA) copolymer.
The basic ideas of the preparation method are that two kinds of mixed anhydride copolymer condensation copolymerizations are made with superficial degradation
Polyestercarbonate acid anhydride copolymer (P (LLA-TMC-SA)).
It is concretely comprised the following steps：
First, terminal hydroxy group PLLA-PTMC copolymers are prepared
With lactide (L-LA) and trimethylene carbonate (TMC) for raw material, ethylene glycol is initiator, and stannous octoate is to urge
Agent, under nitrogen protection, 8h is reacted at 160 DEG C using ring-opening polymerisation method, obtain the PLLA-PTMC that end group is hydroxyl and be copolymerized
Thing.Product is dissolved with dichloromethane complete, add absolute methanol and precipitated, abandoning supernatant, it is viscous to obtain white 3 times repeatedly
Thick shape product (i.e. terminal hydroxy group PLLA-PTMC copolymers), 50 DEG C are dried under vacuum to constant weight.
Secondly, end carboxyl PLLA-PTMC copolymers are prepared
The PLLA-PTMC copolymers of terminal hydroxy group are dissolved in through in dried Isosorbide-5-Nitrae-dioxane, adding simultaneously
Succinic anhydride and DMAP (DMAP), electromagnetic agitation is stayed overnight under nitrogen protection, is synthesized 26h, is used Rotary Evaporators
Unnecessary solvent is removed, concentrate is instilled in excessive cold absolute ether, obtains thick white thing.Thick white thing is dissolved in
In dichloromethane, respectively with the hydrochloric acid and saturated common salt water washing that volume fraction is 10%.Organic phase is dried with anhydrous magnesium sulfate,
Filtering, filtrate is instilled in excessive cold absolute ether, obtain white thick shape product (i.e. end carboxyl PLLA-PTMC copolymers), in
40 DEG C are dried under vacuum to constant weight.
Then, condensation copolymerization prepares P (LLA-TMC-SA)
End carboxyl PLLA-PTMC copolymers and acetic anhydride are flowed back 30 minutes, after cooling, are evaporated under reduced pressure with 50 DEG C of water-baths,
Remove excessive acetic anhydride and the heteroacid of reaction generation；Residual solid is dissolved with anhydrous methylene chloride, removes water petroleum ether precipitation,
Filter, be dried under vacuum to constant weight at room temperature, obtain mixed anhydride prepolymer 1.According to same operation, decanedioic acid (SA) and second are prepared
The mixed anhydride prepolymer 2 of acid anhydrides.
After mixed anhydride prepolymer 1 and mixed anhydride prepolymer 2 are mixed by a certain percentage, melted under the middle and high vacuum of 180 DEG C of oil baths
Melt polyase 13 h, after being cooled to room temperature, dissolved with anhydrous methylene chloride, remove water petroleum ether precipitation, filter, be dried in vacuo at room temperature
To constant weight, polyestercarbonate acid anhydride copolymer (P (LLA-TMC-SA)) is obtained.
Certainly, it will be appreciated by those skilled in the art that those skilled in the art can also refer to the preparation of other prior arts
Obtain the required above-mentioned polyestercarbonate acid anhydride copolymer (P (LLA-TMC-SA) copolymer) of the present invention.The weight that the present invention is paid close attention to
Point is the superficial degradation characteristic using polyestercarbonate acid anhydride copolymer (P (LLA-TMC-SA) copolymer), to realize this hair
Bright technical purpose.
Porcine HGF is a kind of protein molecule that various types of cells can be promoted to breed, and has chemotactic, propagation and reconstruction
Effect, it can induce the target cells such as fibroblast, epithelial cell, vascular endothelial cell in inflammatory phase and 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, hypodermis, bone tissue reparation and blood vessel, nerve reconstructive, promote new vessels generation, nerve cell and bone group
Regeneration is knitted, is served in terms of wound healing considerable.
Porcine HGF has basic fibroblast growth factor, EGF, IGF, nerve
Trophic factors, TGF, keratinocyte growth factor, platelet derived growth factor etc..
Basic fibroblast growth factor promotes cell division propagation and Angiogensis, is to be currently known most strong rush
The Hemapoiesis factor.Fibroblast, epithelial cell, vascular endothelial cell, Skeletal Muscle Cell, cartilage cell, Gegenbaur's cell, 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.
EGF is synthesized by monocyte and ectoderm cell, stimulate the division of internal polytype histocyte and
Propagation, enhancing signaling, merisis and the generation of cytoplasm albumen, such as skin, cornea, lung, tracheae and liver regeneration.
IGF is by two close phases of type-1 insulin like growth factor type and IMA-IGF2BP3-001 type
The family of the small polypeptide composition closed, has and promotes muscle protein synthesis, promotes propagation, the differentiation of sarcoblast, participates in tissue and repair
Multiple and regeneration, so as to accelerate organization healing.
The trace soluble material that neurotrophic factor is present in around sensory neuron, produced by the target cell of neuron
It is raw, specifically promote neure growth and maintenance, promotion organization injury repair.
Porcine HGF is introduced into hydrogel by the present invention, and advantage is in can give in the bio-ink of subsequent configuration
Cells with nutrient.Support is printed with 3D printing technique, after stenter to implant human body, cell factor can promote hydrogel
Cell differentiation, increment on support, accelerate tissue reconstruction.
Sulfa drugs such as sulfapryidine silver, mafenide etc.；QNS 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 have, accelerates blood coagulation
Effect.
In a specific embodiment, the hydrogel can use following component to form：The PLGA- of mass fraction 10%
PEG-PLGA triblock copolymers, the basic fibroblast growth factor of mass fraction 0.01%, the sulphur of mass fraction 0.05%
Amine pyridine silver, surplus are deionized water.
In another specific embodiment, the hydrogel can use following component to form：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 can use following component to form：Mass fraction 20%
PLGA-PEG-PLGA triblock copolymers, the keratinocyte growth factor of mass fraction 0.05%, the peace of mass fraction 0.25%
Network blood, surplus are deionized water.
In an also specific embodiment, the hydrogel can use following component to form：Mass fraction 25%
PLGA-PEG-PLGA triblock copolymers, the IGF of mass fraction 0.5%, the sulfanilamide (SN) of mass fraction 0.3%
Myron and etamcylate, surplus are deionized water.
In an also specific embodiment, the hydrogel can use following component to form：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, present invention also offers superficial degradation type polyestercarbonate acid anhydrides 3D printing biology ink described in example 2
The preparation method of water, comprises the following steps：By the PLGA-PEG-PLGA triblock copolymers of mass fraction 10%~40%, matter
Measure Porcine HGF, the anti-inflammation hemostasia medicine of mass fraction 0.05~0.3% and the going for surplus of fraction 0.001%~1%
Ionized water is blended, and stirs 30min, stands 1~6 hour and the hydrogel is made；The hydrogel and the biological cell is equal
The even gel ink for being mixed into the superficial degradation type polyestercarbonate acid anhydrides 3D printing bio-ink.
Particularly, in Method of printing of the invention, 3D printing gel ink material controls temperature before 3D printing is carried out
Less than phase transition temperature, now gel ink solution is to be in a kind of collosol state of low sticky degree, approximate colloid, passes through addition
Soybean protein isolate solution stirs, and then adds a little calcium sulphate soln, such as add stoste (gel ink) volume 10%
Concentration is 5% soybean protein isolate solution, and the concentration for adding stoste volume 1%-2% is 5-8% calcium sulphate soln, is passed through
Electric charge in sulfuric acid calcium ion with colloid absorption makes colloid trigger cohesion.It is emphasized that this process needs before the printing
15-30 is configured within minute, to prevent coacervation of colloid solidification to be difficult to print.
It needs to be emphasized that step A of the invention 3D printing gel ink material preferably before the printing 15-30
Configure and complete 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 shower nozzle.
In addition, the biological cell suitable for the present invention can be any vertebrate cells, mammalian cell, people
Cell or its combination, its type depend on produced cell construction body, tissue or organ type.Such as the cell can
With including 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, it is 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 addition 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, its 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.Add soy bean proteinous soln and
During calcium sulphate soln, it is necessary to maintain the temperature in transition temperature range and carry out.It is follow-up mixed because hydrogel has temperature-responsive
Closing the bio-ink formed 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 standby 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 according to step B afterwards.
Step B：The temperature of the gel ink sump of 3D biometric print machines is controlled to be less than phase transition temperature, printing table top control temperature
Spend for 37 degrees Celsius, using the temperature-responsive of gel ink, by 3D printing shower nozzle by the 3D printing gel ink material
It is printed as model.
In this step, 3D printer can be suitable for the printer of organism 3D printing using existing any one, wherein
Foregoing standby 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 stably easy to print and be not easily blocked shower nozzle, and the 3D printing shower nozzle of printer can set and be located at
In temperature control box, and printing forms model on the printing table top in temperature control box.Now, control the temperature in temperature control box Celsius for 37
Degree so that model being capable of solidifying and setting.
If for the 3D printing bio-ink in aforementioned exemplary 2, used 3D printer can use existing
It is suitable for the 3D printer with two coaxially arranged shower nozzles of organism 3D printing, the 3D printer has a gel ink
Sump and a support ink storehouse, printing step and abovementioned steps B principle 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, using coaxially arranged with two
The 3D printer of shower nozzle, support is formed by support ink described in the inner circumferential side nozzle printing of the 3D printer, by described
The 3D printing gel ink material is printed as model by the outer circumferential side shower nozzle of 3D printer around the support.Certainly, print
During, gel ink is the nozzle printing being arranged on the outside of coaxitron by 3D printer, and support ink is to pass through 3D
The nozzle printing being arranged on the inside of coaxitron of printer.
It should be noted that soybean protein isolate is with the addition of in abovementioned steps is molten for gel ink specifically preferred according to the invention
Liquid and calcium sulphate soln have carried out pre-agglomeration, condense solidification after being printed as model.The change and the regulation and control of time of temperature make biology
Ink possesses two kinds of coacervation processes, and when being printed to by printing head on temperature control box printing table top, the droplet printed every time is very
Fixed structure is easily formed, thus the above-mentioned particularly preferred gel ink of the present invention can need not print one layer and cover afterwards
One layer of timbering material is supported, there is provided one kind is without support 3D biometric print technologies.Namely pass through the above-mentioned 3D of the present invention
The step A and step B of Method of printing, the 3D printing model without support support can be directly obtained on the basis of hydrogel, can
To save the time-write interval of support, without changing printhead, can save time-write interval and cost.
Certainly, be not precluded from can be by existing 3D printer successively printing alternate bio-ink and support material by the present invention
Material forms the technical scheme of model, i.e. in order to form the model structure of complexity, in a preferred embodiment, this step enters one
Step is included in the bio-ink and the timbering material successively printing alternate into model in temperature control box by 3D printing shower nozzle,
To provide support to bio-ink by timbering material.Wherein, the timbering material can be selected from any existing suitable
For the timbering material of 3D printing, including it is not limited to fibrin, alginates, agarose, chitosan and combinations thereof.
Or as described in aforementioned exemplary 2, gel ink of the invention can also pass through the 3D printing of coaxially arranged shower nozzle
It is support that machine, which is obtained with the inner circumferential side of support ink cooperation, the structural model that outer circumferential side is gel growth matrix.
Wherein, the preparation of the gel ink of outer circumferential side and print conditions are identical with the print procedure of no rack structure model,
The melting temperature of the support ink of inner circumferential side, such as polyestercarbonate acid anhydride copolymer (P (LLA-TMC-SA) copolymer) are molten
Melt temperature as more than 100 degrees Celsius, therefore, support ink storehouse needs to keep the condition of high temperature when printing, logical after printing
Cross low temperature and be formed by curing support.The gel ink and support ink that the example 2 of the present invention uses possess the two kinds of phases completely contradicted
Transition process, gel ink are needed to be printed with liquid under low-temperature condition, solidified by 37 degrees Celsius of table top temperature；And
Support ink needs to form printable flow-like under the condition of high temperature more than 100 degrees Celsius, while in 37 degrees Celsius of platform
Face temperature is solidified.And coaxitron gradient printing type of the present invention, can be when two shower nozzles work, profit
The low temperature gel ink for accelerating outside with the high temperature support ink of inner side solidifies, while utilizes the low temperature gel ink in outside to accelerate
The solidification of the high temperature support ink of inner side.Thus, the 3D printing ink of example 2 of the invention can substantially reduce table top temperature
Control accuracy, faster, printing effect is also higher for curing rate.
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 treatment, 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 omit by
Model inserts the step of temperature control box), the temperature in the temperature control box is slowly reduced to less than 0 degree, makes the moisture in the model
Freeze completely.The effect of this step is 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 dispersed to congeal into ice, and keeps model as far as possible for water in model
Internal structure, the too fast meeting that otherwise cools cause model overall structure to destroy, are difficult to keep tissue morphology after ice-out.It is preferred that
In this step, 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 cause biological cell loss of activity.Therefore, to carry out
Step C refrigerating process is, it is necessary in foregoing step A, before biological cell is prepared into 3D printing bio-ink, to life
Thing cell carries out protection processing：The starch solution that the glycerine and concentration for being 10% with concentration by biological cell are 10% uniformly mixes
Close and stand 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 step C slow freezing, ICW appears, and reduces ice crystal and is formed,
So as to avoid cellular damage, ensure biological cell survival.In a specific embodiment, the glycerine of concentration 10% and concentration are
The addition 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 also cause model to cave in.Bio-ink of the present invention has two kinds and condensed
Journey, it is a kind of be 3D printing gel ink material temperature-responsive cohesion, it is a kind of be add after soybean protein and calcium sulfate when
Between control condense.In temperature-fall period, moisture can be very good to seek connections with soybean protein and sulphur 3D biomaterials before freezing
Sour calcium is condensed on the skeleton to be formed, and is unlikely to cave in.
Step D：The temperature in the temperature control box is quickly raised to 37 degrees Celsius, makes the hydrogel in the model completely solid
Change, so as 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 generally little, the water in model all condenses, now can be by being rapidly heated
To 37 degree of organism active temperature, the hydrogel in model is set to solidify again, while the ice-out in model, in gel solidification
During moisture deviate from, the hole containing ice can remain in the gel structure of solidification, so as to form containing biological cell
Loose structure.
Particularly, the temperature-rise period in this step is different from foregoing temperature-fall period, it is necessary to which lift scheme temperature makes as early as possible
Obtain structure as early as possible to solidify, avoid solidification from slowly causing hole to disappear.In a preferred embodiment, can be to institute in this step
The dry-heat air of 37 degree of conveying in temperature control box is stated, the ice rapid translating in the model is escaped into gas from the model, from
And form loose structure on the model.Dry-heat air can lift temperature in temperature control box quickly to 37 degree, dry simultaneously
Hot blast has discharged the moisture in temperature control box, reduces water vapor partial pressure, has further speeded up the speed that moisture is converted into gas, thus
The present embodiment can reach the effect for making model be formed by curing loose structure as early as possible.
After the loose structure containing biological cell obtained on model, model can be placed in nutritive solution and carried out
Culture, because loose structure is that biological cell provides and obtains the passage of nutrition and metabolism, thus cell inside model is the same as outside
Portion's cell equally can obtain enough nutrient growth, can be obtained by this 3D printing method of the present invention bigger thicker
Institutional framework.
In order to avoid institutional framework growth closure after, internal cell easily lack nutrient, metabolism it is not smooth and dead, another
In one preferred embodiment, after the porous structural model containing biological cell is obtained, injected in the loose structure
Vascular endothelial cell, vascular endothelial cell is attached on structure pore framework, model is placed in nutritive solution carried out afterwards
Culture, goes out blood vessel, original cell growth is into required tissue in model, the blood vessel grown by vascular endothelial cell growth
Further nutrient and metabolic pathway are provided for required tissue.Further, in order to avoid the hole inside loose structure does not connect
It is logical, the defects of causing the blood vessel of growth to be difficult to insertion, 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 insertion by laser, for by the hole in model
Hole connection is got up.Laser boring obtain through-hole diameter it is limited, be appropriate only for by hole connection be easy to vascular endothelial cell and
Nutrient solution enters inner void, is operated according to aforesaid way thereafter, vascular endothelial cell is injected in loose structure, is made intravascular
Chrotoplast is attached on structure pore framework, model is placed in nutritive solution cultivated afterwards, pass through vascular endothelial cell
Grow blood vessel.With the growth of blood vessel and tissue, the through hole for these insertions 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 includes an independent technical scheme.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 scheme involved in each embodiment
The modes of different embodiments can be mutually combined into understand protection scope of the present invention by regarding as.
The schematical embodiment of the present invention is the foregoing is only, is not limited to the scope of the present invention.It is any
Those skilled in the art, equivalent variations, modification and the combination made on the premise of the design of the present invention and principle is not departed from,
The scope of protection of the invention all should be belonged to.
Claims (7)
- A kind of a kind of a kind of 1. superficial degradation type polyestercarbonate acid anhydrides 3D printing bio-ink, by gel ink and support ink Form, the gel ink is made up of hydrogel and biological cell, it is characterised in that the hydrogel is made up of following component： The PLGA-PEG-PLGA triblock copolymers of mass fraction 10%~40%, the cell growth of mass fraction 0.001%~1% The factor, the anti-inflammation hemostasia medicine of mass fraction 0.05~0.3%, surplus are deionized water；The support ink is by the gel The polyestercarbonate acid anhydride copolymer of the 5%~50% of the gross mass of ink is formed.
- 2. a kind of preparation method of superficial degradation type polyestercarbonate acid anhydrides 3D printing bio-ink as claimed in claim 1, Comprise the following steps：By the PLGA-PEG-PLGA triblock copolymers of mass fraction 10%~40%, mass fraction 0.001% The deionized water blending of~1% Porcine HGF, the anti-inflammation hemostasia medicine of mass fraction 0.05~0.3% and surplus, is stirred 30min is mixed, 1~6 hour is stood and the hydrogel is made；The hydrogel and the biological cell are uniformly mixed into the table The gel ink of face degradation-type polyestercarbonate acid anhydrides 3D printing bio-ink.
- A kind of 3. 3D printing side of superficial degradation type polyestercarbonate acid anhydrides 3D printing bio-ink using described in claim 1 Method, comprise the following steps：Step A：Using the superficial degradation type polyestercarbonate acid anhydrides 3D printing bio-ink described in claim 1, the gel Ink adds soybean protein isolate solution and stirred, then add calcium sulphate soln under the temperature conditionss less than phase in version It is uniformly mixed into standby 3D printing gel ink material；Step B：The temperature of the gel ink sump of 3D biometric print machines is controlled to be less than phase transition temperature, printing table top temperature is taken the photograph for 37 Family name's degree, using the 3D printer with two coaxially arranged shower nozzles, pass through the inner circumferential side nozzle printing institute of the 3D printer State support ink and form support, the 3D printing gel ink material is surrounded by institute by the outer circumferential side shower nozzle of the 3D printer State support and be printed as model.
- 4. method as claimed in claim 3, it is characterised in that in the step A, the biological cell is beaten preparing the 3D Before printing bio-ink, protection processing is carried out to the biological cell：By glycerine that the biological cell and concentration are 10% with And the starch solution that concentration is 10% uniformly mixes and stands 2-3 hours.
- 5. method as claimed in claim 4, further comprises following steps：Step C：The model is placed in temperature control box, the temperature in the temperature control box is slowly reduced to less than 0 degree, makes the mould Moisture in type freezes completely；Step D：The temperature in the temperature control box is quickly raised to 37 degrees Celsius, the hydrogel in the model is fully cured, So as to obtain the porous structural model containing biological cell.
- 6. 3D printing method as claimed in claim 5, it is characterised 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.
- 7. 3D printing method as claimed in claim 5, it is characterised in that in the step D, 37 are conveyed into the temperature control box The dry-heat air of degree, the ice rapid translating in the model is set to be escaped into gas from the model, so as to the shape on the model Into loose 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 |
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| CN113180879A (en) * | 2021-05-24 | 2021-07-30 | 上海大学 | Biological tissue preparation system and method |
| CN113180879B (en) * | 2021-05-24 | 2023-12-22 | 上海大学 | Biological tissue preparation system and method |
| CN114259604A (en) * | 2021-12-17 | 2022-04-01 | 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 |
| EP4349778A1 (en) * | 2022-10-03 | 2024-04-10 | Universitat Internacional De Catalunya, Fundació Privada | Ink for 3d printing |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107523136A (en) | 2017-12-29 |
| CN107523136B (en) | 2020-01-14 |
| CN107400412B (en) | 2018-08-24 |
| CN107502061B (en) | 2018-08-24 |
| CN107502061A (en) | 2017-12-22 |
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