CN106378930B - The photocuring 3D printing method of temperature sensitive culture surface material - Google Patents
The photocuring 3D printing method of temperature sensitive culture surface material Download PDFInfo
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- CN106378930B CN106378930B CN201610834643.5A CN201610834643A CN106378930B CN 106378930 B CN106378930 B CN 106378930B CN 201610834643 A CN201610834643 A CN 201610834643A CN 106378930 B CN106378930 B CN 106378930B
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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
- B33Y30/00—Apparatus for additive manufacturing; Details thereof or accessories therefor
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B08—CLEANING
- B08B—CLEANING IN GENERAL; PREVENTION OF FOULING IN GENERAL
- B08B3/00—Cleaning by methods involving the use or presence of liquid or steam
- B08B3/04—Cleaning involving contact with liquid
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B08—CLEANING
- B08B—CLEANING IN GENERAL; PREVENTION OF FOULING IN GENERAL
- B08B3/00—Cleaning by methods involving the use or presence of liquid or steam
- B08B3/04—Cleaning involving contact with liquid
- B08B3/10—Cleaning involving contact with liquid with additional treatment of the liquid or of the object being cleaned, e.g. by heat, by electricity or by vibration
- B08B3/12—Cleaning involving contact with liquid with additional treatment of the liquid or of the object being cleaned, e.g. by heat, by electricity or by vibration by sonic or ultrasonic vibrations
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- Engineering & Computer Science (AREA)
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
Abstract
The present invention discloses a kind of photocuring 3D printing method of temperature sensitive culture surface material, includes the following steps：S1）Prepare photocuring 3D printing temperature sensing material；S2）Temperature sensitive printed material jet printing on substrate surface and is carried out by ultraviolet or laser irradiation grafting curing reaction using photocuring 3D printer；S3）Make itself and the further graft curing of base material in band type heated at constant temperature channel；S4）The temperature sensitive culture surface product utilization ultra-pure water of preparation is cleaned, is dried.The obtained temperature sensitive culture surface thickness of the present invention more uniformity, the quality control for product of being more convenient for, equipment cost is cheap, and prepare with scale reproducibility is more preferable, it is easier to realize.
Description
Technical field
The present invention relates to photocuring 3D printing technique fields, are to be related to a kind of temperature sensitive culture surface material more specifically
Photocuring 3D printing preparation method.
Background technology
With being constantly progressive for mankind's scientific and technological level, the achievement in temperature-responsive investigation of materials field also becomes increasingly abundant.Its
In, temperature-responsive material is applied to the research opinion in the fields such as cell and tissue culture, medicament slow release, Protein Separation and purifying
Text and patent report are also increasing in nearest decades.In biology, drug and medical domain, these temperature-responsive materials
The middle research report for belonging to n-isopropyl acrylamide class temperature sensitive polymer is most.This kind of temperature sensing material becomes there are one structure with temperature
Change and changed characteristic, changes so as to cause the hydrophilic-hydrophobic performance of material.There are one face the transformation of its hydrophilic and hydrophobic
Boundary's transition temperature（LCST）, it is higher than this temperature, material shows as hydrophobic；Less than this temperature, material shows as hydrophilic.Thin
Born of the same parents' biology and organizational engineering field are sent out using n-isopropyl acrylamide class temperature sensitive polymer with the variation of environment temperature
The characteristic of raw hydrophilic and hydrophobic sex reversal can realize the not damaged receipts of cell under conditions of without outer addition proteolytic enzyme
It obtains or the not damaged transfer of tissue lamellar, toxigenic capacity and raising to reduce cell and tissue lamellar harvests cell and tissue
The perfect degree and function of lamella.
In the document and patent reported at present, the preparation method on this temperature sensitive surface for being used for carrying out cell and tissue culture
In, it is most of be all using n-isopropyl acrylamide class monomer is first configured to certain density temperature sensitive solution, or then
The temperature sensitive solution is immersed in temperature sensitive liquid using hardened coating in the substrate surface of graft polymerization preformer or by the substrate surface of graft polymerization preformer
In further take out after a certain period of time, finally carry out high energy electron beams carry out graft reaction.Temperature sensitive culture surface is prepared in this
There are many disadvantages for method, and being that high energy electron beam device is extremely expensive first makes it be difficult to popularize, and also hardened coating can make temperature
Quick substance distribution makes this method poor reproducibility in uneven thickness, furthermore the electron beam irradiation of high dose can make the base material of graft polymerization preformer
Color change occurs, mechanical property reduces and final product performance is made to be deteriorated.
In existing patent and document report, also there is seldom researcher using photosensitizer is added in temperature sensitive liquid, so
The temperature sensitive liquid coating for carrying out substrate surface afterwards, finally prepares temperature sensitive culture surface with UV illumination initiation grafting polymerisation
Method.This method is equally the presence of temperature sensitive polymer layer distribution and in uneven thickness, and by single ultraviolet
Light radiation polymerization mode, temperature sensitive monomer are difficult that reaction is thorough.In addition, some researcher is first in the base material table of graft polymerization preformer
Face introduces the group of halogen atom, and then causing temperature sensitive monomer using atom transfer radical polymerization principle carries out fixed point polymerization,
To realize the temperature-sensing property of culture surface.The preparation method of this kind of temperature sensitive culture surface although finally prepd temperature sensitive superficiality
Can be than more uniform, but technics comparing is complicated, manufacturing cost is higher, and grafting amount is relatively low, is prepared so being difficult to realize batch.
3D printing technique is to first pass through Computerized three-dimensional software to carry out three-dimensional graphics design to pre-prepared product, then will
3-D graphic information is converted into 3D printing code and is entered into 3D printer, is superimposed most by the printing layer by layer of printer
The intelligent Manufacturing Technology of entity is formed afterwards.In all 3D printing techniques, photocuring 3D printing technique manufactures fastest,
And printing precision is high, and favorable reproducibility, thickness error can be controlled in positive and negative 0.001~0.002mm, be one kind most useful for scale
The intelligent Manufacturing Technology of change.But photocuring 3D printing technique is applied in the preparation of temperature sensitive culture surface and yet there are no document
And patent report.
Invention content
The purpose of the present invention is to provide a kind of photocuring 3D printing preparation methods of temperature sensitive culture surface material, to overcome
Deficiency in the presence of the prior art.
The present invention the technical solution to solve the technical problem is that：A kind of photocuring 3D printing side of temperature sensitive culture surface material
Method includes the following steps：S1）Prepare photocuring 3D printing temperature sensing material；S2）Using photocuring 3D printer by temperature sensitive printing material
Expect jet printing on substrate surface and carries out ultraviolet or laser irradiation grafting curing reaction；S3）In band type heated at constant temperature
Make itself and the further graft curing of base material in channel；S4）The temperature sensitive culture surface product utilization ultra-pure water of preparation is cleaned,
Drying.
The substrate surface, which refers to that surface is treated, can be condensed containing abundant hydroxyl or carboxyl and with silane
The glass of reaction, ceramics or plastics surface.
The photocuring 3D printer refers to Ployjet class ejection-type photocuring 3D printers, and the light source of radiation curing is adopted
With ultraviolet or laser light source, the layer thickness control of cured temperature sensitive polymer film is printed between 0.01~0.05mm, it is vertical to print
Speed control is in 0.1~0.3mm/min.
In S4 steps, temperature sensitive culture surface product is using soaking and washing and the processing of the combined type cleaning of ultrasonic cleaning or list
Solely using ultrasonic cleaning.
The soaking and washing time is 8~72h.
The concussion frequency of ultrasonic cleaning is 20~150KHz, and the time is 5~60min.
The photocuring 3D printing temperature sensing material includes temperature sensitive monomer, grafted monomers, crosslinking agent, chemical initiator, photosensitive
Initiator and solvent, wherein the molar ratio of temperature sensitive monomer and grafted monomers is 95:5~85:In 15 ranges, crosslinking agent is temperature
The 0.5%~1% of quick monomer and grafted monomers total moles quantity, chemical initiator are temperature sensitive monomer and grafted monomers total moles quantity
0.5%~1%, photoinitiator is the 0.5%~2% of temperature sensitive monomer and grafted monomers total moles quantity.
The temperature sensitive monomer includes n-isopropyl acrylamide or N- isopropyl -2- Methacrylamides or N- isopropyls
The blend or n-isopropyl acrylamide and N- isopropyl -2- methyl of acrylamide and N- isopropyl -2- Methacrylamides
The oligomer of acrylamide, or by n-isopropyl acrylamide of the molecular weight less than 2000 and N- isopropyl -2- methacryls
The blend of the oligomer mixing gained of amine.
The grafted monomers include the silane coupling agent containing unsaturated double-bond, allyltriethoxysilane, allyl
Base trimethoxy silane, 2- methacrylics triethoxysilane, 2- methacrylics trimethoxy silane, propyl acrylate-
3- trimethoxy silanes, propyl acrylate -3- triethoxysilanes, propyl methacrylate -3- trimethoxy silanes, methyl-prop
The one of which of olefin(e) acid propyl ester -3- triethoxysilanes or arbitrary two kinds or two or more of mixture.
The crosslinking agent includes diacrylate -1,4-butanediol ester, ethylene glycol dimethacrylate, N, N- di-2-ethylhexylphosphine oxides
Acrylimide, bismaleimide amine substance, the crosslinking agent contain two or more unsaturated double-bonds；The chemistry
Initiator includes azodiisobutyronitrile, azobisisoheptonitrile or di-t-butyl peroxide, benzoyl peroxide, peroxidating diisopropyl
Benzene, potassium peroxydisulfate, ammonium persulfate；The photoinitiator using can directly decompose under light illumination production free radical or anion,
The benzoin and its derivatives or acetophenones derivative or benzophenone and its derivative of cation；The solvent is polarity
Aprotic solvent, including n,N-Dimethylformamide, n,N-dimethylacetamide, N-Methyl pyrrolidone, tetrahydrofuran, ring
The one or more of which of hexanone, the solvent are the low toxicity compounds that do not reacted with hydroxyl and carboxyl.
The beneficial effects of the invention are as follows：
The principle of the present invention is will prepared temperature sensing material combination in advance using photocuring 3D printing intelligent Manufacturing Technology
Object is first injected in the substrate surface of graft polymerization preformer, then causes temperature sensitive rmc monomer graft curing with the irradiation of ultraviolet or laser, then
Heating further carries out graft curing reaction, keeps its reaction more thorough, and temperature sensitive polymer graft reaction is more perfect, finally carries out clear
Washing may remaining small-molecule substance to remove temperature sensitive culture surface.
The beneficial effects of the invention are as follows the temperature sensitive culture surface thickness prepared using photocuring 3D printing technique is more uniform
Unanimously, it is more convenient for product compared with then the temperature sensitive substance of the hardened coating of existing literature and patent report carries out electron beam irradiation
Quality control, equipment cost is cheap, and prepare with scale reproducibility is more preferable, it is easier to realize.Therefore, the present invention prepares temperature sensitive culture
The market prospects of surface product will be more wide.
Description of the drawings
Fig. 1 is the temperature sensitive culture surface material 3T3 cell culture pictures of 3D printing.
Fig. 2 is that the temperature sensitive culture surface material of 3D printing carries out picture of the 3T3 cell culture tests after cell detachment.
Specific implementation mode
The embodiment of the present invention is described below in detail, it is noted that embodiment described in the invention is exemplary, only
For explaining the present invention, and it is not considered as limiting the invention.
Embodiment 1
The present embodiment prepares a kind of temperature sensitive culture surface product, and detailed preparation process is as follows：
According to molar ratio 95:5 weigh n-isopropyl acrylamide and allyltriethoxysilane, then weigh and rub respectively
Your number is 0.5% N of the sum of the above two, and N- methylene-bisacrylamides, benzoyl peroxide, styrax, it is total to be eventually adding
The tetrahydrofuran of mass fraction 40% makes it fully dissolve as solvent, stirring, is filtered with 300 mesh screens for use.
Silicate glass platelet substrate material is impregnated in sodium hydrate aqueous solution and hydrogen peroxide aqueous sulfuric acid respectively
Processing, drying.The prepared temperature sensing material mixed solution in front is added to the resin liquid pool of ejection-type photocuring 3D printer
In, input the print code got well and converted with computer 3D Software for Design in advance（The temperature sensitive thickness of design printing is in this embodiment
10um）.Previously processed glass flake base material is placed on the printing bed of 3D printer, 3D printer is opened and starts to beat
Print.After waiting for 3D printer printing, the glass flake for printing temperature sensitive layer is put into 100 DEG C of heated constant temperature channel at heating
Reason 20min makes its further graft curing.After having handled, soaking and washing 48h in RO water is put it into, is surpassed under 50KHz frequencies
Sound cleans 5 minutes, and it is 68 in 25 DEG C of contact angle then to dry the quick culture surface of thermometric with watero, and send to gamma radiation disinfection
3T3 cell culture tests are carried out afterwards.Cell culture condition is：5% CO2, contain 10% fetal calf serum basal medium, 37 DEG C
48h is cultivated in incubator（As shown in Figure 1）, 20min then is placed under the conditions of 20 DEG C, under gently blowing and beating 15 with pasteur pipet,
Cell can all fall off in flakes（As shown in Figure 2）.
Embodiment 2
The present embodiment prepares a kind of temperature sensitive culture surface product, and detailed preparation process is as follows：
According to molar ratio 95:5 weigh n-isopropyl acrylamide and allyltriethoxysilane, then weigh and rub respectively
Your number is 0.8% N of the sum of the above two, and N- methylene-bisacrylamides, benzoyl peroxide, styrax, it is total to be eventually adding
The tetrahydrofuran of mass fraction 40% makes it fully dissolve as solvent, stirring, is filtered with 300 mesh screens for use.
Silicate glass platelet substrate material is impregnated in sodium hydrate aqueous solution and hydrogen peroxide aqueous sulfuric acid respectively
Processing, drying.The prepared temperature sensing material mixed solution in front is added to the resin liquid pool of ejection-type photocuring 3D printer
In, input the print code got well and converted with computer 3D Software for Design in advance（The temperature sensitive thickness of design printing is in this embodiment
10um）.Previously processed glass flake base material is placed on the printing bed of 3D printer, 3D printer is opened and starts to beat
Print.After waiting for 3D printer printing, the glass flake for printing temperature sensitive layer is put into 100 DEG C of heated constant temperature channel at heating
Reason 20min makes its further graft curing.After having handled, soaking and washing 48h in RO water is put it into, is surpassed under 50KHz frequencies
Sound cleans 5 minutes, and it is 70 in 25 DEG C of contact angle then to dry the quick culture surface of thermometric with watero, and send to gamma radiation disinfection
3T3 cell culture tests are carried out afterwards.Cell culture condition is：5% CO2, contain 10% fetal calf serum basal medium, 37 DEG C
20min is placed under the conditions of 20 DEG C after cultivating 48h in incubator, under gently blowing and beating 13 with pasteur pipet, cell can be complete in flakes
Portion falls off.
Embodiment 3
The present embodiment prepares a kind of temperature sensitive culture surface product, and detailed preparation process is as follows：
According to molar ratio 90:10 weigh n-isopropyl acrylamide and allyltriethoxysilane, then weigh respectively
Molal quantity be the above two the sum of 0.8% N, N- methylene-bisacrylamides, benzoyl peroxide, styrax, be eventually adding for
The tetrahydrofuran of total mass fraction 40% makes it fully dissolve as solvent, stirring, is filtered with 300 mesh screens for use.
Silicate glass platelet substrate material is impregnated in sodium hydrate aqueous solution and hydrogen peroxide aqueous sulfuric acid respectively
Processing, drying.The prepared temperature sensing material mixed solution in front is added to the resin liquid pool of ejection-type photocuring 3D printer
In, input the print code got well and converted with computer 3D Software for Design in advance（The temperature sensitive thickness of design printing is in this embodiment
10um）.Previously processed glass flake base material is placed on the printing bed of 3D printer, 3D printer is opened and starts to beat
Print.After waiting for 3D printer printing, the glass flake for printing temperature sensitive layer is put into 100 DEG C of heated constant temperature channel at heating
Reason 20min makes its further graft curing.After having handled, soaking and washing 48h in RO water is put it into, is surpassed under 50KHz frequencies
Sound cleans 5 minutes, and it is 72 in 25 DEG C of contact angle then to dry the quick culture surface of thermometric with watero, and send to gamma radiation disinfection
3T3 cell culture tests are carried out afterwards.Cell culture condition is：5% CO2, contain 10% fetal calf serum basal medium, 37 DEG C
20min is placed under the conditions of 20 DEG C after cultivating 48h in incubator, under gently blowing and beating 11 with pasteur pipet, cell can be complete in flakes
Portion falls off.
Embodiment 4
The present embodiment prepares a kind of temperature sensitive culture surface product, and detailed preparation process is as follows：
According to molar ratio 90:10 weigh n-isopropyl acrylamide and allyltriethoxysilane, then weigh respectively
Molal quantity is 0.8% N of the sum of the above two, and N- methylene-bisacrylamides, azodiisobutyronitrile and styrax are eventually adding
For total mass fraction 50% tetrahydrofuran as solvent, stirring makes it fully dissolve, and is filtered with 300 mesh screens for use.
Silicate glass platelet substrate material is impregnated in sodium hydrate aqueous solution and hydrogen peroxide aqueous sulfuric acid respectively
Processing, drying.The prepared temperature sensing material mixed solution in front is added to the resin liquid pool of ejection-type photocuring 3D printer
In, input the print code got well and converted with computer 3D Software for Design in advance（The temperature sensitive thickness of design printing is in this embodiment
10um）.Previously processed glass flake base material is placed on the printing bed of 3D printer, 3D printer is opened and starts to beat
Print.After waiting for 3D printer printing, the glass flake for printing temperature sensitive layer is put into 100 DEG C of heated constant temperature channel at heating
Reason 20min makes its further graft curing.After having handled, soaking and washing 48h in RO water is put it into, is surpassed under 50KHz frequencies
Sound cleans 5 minutes, and it is 75 in 25 DEG C of contact angle then to dry the quick culture surface of thermometric with watero, and send to gamma radiation disinfection
3T3 cell culture tests are carried out afterwards.Cell culture condition is：5% CO2, contain 10% fetal calf serum basal medium, 37 DEG C
20min is placed under the conditions of 20 DEG C after cultivating 48h in incubator, under gently blowing and beating 15 with pasteur pipet, cell can be complete in flakes
Portion falls off.
Embodiment 5
The present embodiment prepares a kind of temperature sensitive culture surface product, and detailed preparation process is as follows：
According to molar ratio 90:10 weigh n-isopropyl acrylamide and allyltriethoxysilane, then weigh respectively
Molal quantity is 0.8% N of the sum of the above two, and N- methylene-bisacrylamides, azodiisobutyronitrile and styrax are eventually adding
For total mass fraction 50% tetrahydrofuran as solvent, stirring makes it fully dissolve, and is filtered with 300 mesh screens for use.
Silicate glass platelet substrate material is impregnated in sodium hydrate aqueous solution and hydrogen peroxide aqueous sulfuric acid respectively
Processing, drying.The prepared temperature sensing material mixed solution in front is added to the resin liquid pool of ejection-type photocuring 3D printer
In, input the print code got well and converted with computer 3D Software for Design in advance（The temperature sensitive thickness of design printing is in this embodiment
15um）.Previously processed glass flake base material is placed on the printing bed of 3D printer, 3D printer is opened and starts to beat
Print.After waiting for 3D printer printing, the glass flake for printing temperature sensitive layer is put into 100 DEG C of heated constant temperature channel at heating
Reason 20min makes its further graft curing.After having handled, soaking and washing 48h in RO water is put it into, is surpassed under 50KHz frequencies
Sound cleans 5 minutes, and it is 65 in 25 DEG C of contact angle then to dry the quick culture surface of thermometric with watero, and send to gamma radiation disinfection
3T3 cell culture tests are carried out afterwards.Cell culture condition is：5% CO2, contain 10% fetal calf serum basal medium, 37 DEG C
20min is placed under the conditions of 20 DEG C after cultivating 48h in incubator, under gently blowing and beating 10 with pasteur pipet, cell can be complete in flakes
Portion falls off.
Embodiment 6
The present embodiment prepares a kind of temperature sensitive culture surface product, and detailed preparation process is as follows：
According to molar ratio 90:10 weigh n-isopropyl acrylamide and allyltriethoxysilane, then weigh respectively
Molal quantity is 0.8% N of the sum of the above two, and N- methylene-bisacrylamides, azodiisobutyronitrile and styrax are eventually adding
For total mass fraction 50% tetrahydrofuran as solvent, stirring makes it fully dissolve, and is filtered with 300 mesh screens for use.
Silicate glass platelet substrate material is impregnated in sodium hydrate aqueous solution and hydrogen peroxide aqueous sulfuric acid respectively
Processing, drying.The prepared temperature sensing material mixed solution in front is added to the resin liquid pool of ejection-type photocuring 3D printer
In, input the print code got well and converted with computer 3D Software for Design in advance（The temperature sensitive thickness of design printing is in this embodiment
15um）.Previously processed glass flake base material is placed on the printing bed of 3D printer, 3D printer is opened and starts to beat
Print.After waiting for 3D printer printing, the glass flake for printing temperature sensitive layer is put into 100 DEG C of heated constant temperature channel at heating
Reason 30min makes its further graft curing.After having handled, soaking and washing 48h in RO water is put it into, is surpassed under 50KHz frequencies
Sound cleans 5 minutes, and it is 68 in 25 DEG C of contact angle then to dry the quick culture surface of thermometric with watero, and send to gamma radiation disinfection
3T3 cell culture tests are carried out afterwards.Cell culture condition is：5% CO2, contain 10% fetal calf serum basal medium, 37 DEG C
20min is placed under the conditions of 20 DEG C after cultivating 48h in incubator, under gently blowing and beating 11 with pasteur pipet, cell can be complete in flakes
Portion falls off.
Embodiment 7
The present embodiment prepares a kind of temperature sensitive culture surface product, and detailed preparation process is as follows：
According to molar ratio 90:10 weigh n-isopropyl acrylamide and allyltriethoxysilane, then weigh respectively
Molal quantity is 0.8% N of the sum of the above two, and N- methylene-bisacrylamides, azodiisobutyronitrile and styrax are eventually adding
For total mass fraction 50% tetrahydrofuran as solvent, stirring makes it fully dissolve, and is filtered with 300 mesh screens for use.
Silicate glass platelet substrate material is impregnated in sodium hydrate aqueous solution and hydrogen peroxide aqueous sulfuric acid respectively
Processing, drying.The prepared temperature sensing material mixed solution in front is added to the resin liquid pool of ejection-type photocuring 3D printer
In, input the print code got well and converted with computer 3D Software for Design in advance（The temperature sensitive thickness of design printing is in this embodiment
10um）.Previously processed glass flake base material is placed on the printing bed of 3D printer, 3D printer is opened and starts to beat
Print.After waiting for 3D printer printing, the glass flake for printing temperature sensitive layer is put into 100 DEG C of heated constant temperature channel at heating
Reason 30min makes its further graft curing.After having handled, soaking and washing 48h in RO water is put it into, is surpassed under 50KHz frequencies
Sound cleans 10 minutes, and it is 74 in 25 DEG C of contact angle then to dry the quick culture surface of thermometric with watero, and send to gamma radiation disinfection
3T3 cell culture tests are carried out afterwards.Cell culture condition is：5% CO2, contain 10% fetal calf serum basal medium, 37 DEG C
20min is placed under the conditions of 20 DEG C after cultivating 48h in incubator, under gently blowing and beating 13 with pasteur pipet, cell can be complete in flakes
Portion falls off.
Embodiment 8
The present embodiment prepares a kind of temperature sensitive culture surface product, and detailed preparation process is as follows：
According to molar ratio 90:10 weigh n-isopropyl acrylamide and allyl -3- triethoxysilanes, then claim respectively
0.8% N that molal quantity is the sum of the above two is measured, N- methylene-bisacrylamides, azodiisobutyronitrile, styrax are eventually adding
For total mass fraction 50% tetrahydrofuran as solvent, stirring makes it fully dissolve, and is filtered with 300 mesh screens for use.
Plastic substrate is subjected to surface plasma hydrophilic treated, introduces hydroxyl and carboxylic group in substrate surface, so
The prepared temperature sensing material mixed solution in front is added in the resin liquid pool of ejection-type photocuring 3D printer afterwards, input is pre-
The print code first got well and converted with computer 3D Software for Design（It is 10um that design, which prints temperature sensitive thickness, in this embodiment）.Will before
The plastic basis material that surface treatment is crossed is placed on the printing bed of 3D printer, is opened 3D printer and is started to print.Wait for 3D printer
After printing, the plastic tab for printing temperature sensitive layer is put into 85 DEG C of heated constant temperature channel heat 30min make its into
One step graft curing.After having handled, soaking and washing 48h in RO water is put it into, is cleaned by ultrasonic 5 minutes under 50KHz frequencies, so
The drying quick culture surface of thermometric is 70 in 25 DEG C of contact angle with water afterwardso, and carry out 3T3 cell trainings after sending to gamma radiation disinfection
Support test.Cell culture condition is：5% CO2, contain the basal medium of 10% fetal calf serum, 37 DEG C are cultivated 48h in the incubator
20min is placed under the conditions of 20 DEG C afterwards, under gently blowing and beating 11 with pasteur pipet, cell can all fall off in flakes.
It need not added by the cell of the temperature sensitive culture surface culture prepared under the conditions of implementation 1 to 8 various combination of embodiment
Under conditions of adding pancreatin to digest, the harvest of cell can be realized well at low temperature, it was demonstrated that the present invention can very efficiently
Prepare in homogeneous thickness, the good temperature sensitive culture surface of temperature-responsive.
Although an embodiment of the present invention has been shown and described, it will be understood by those of ordinary skill in the art that：Not
In the case of being detached from the principle of the present invention and objective a variety of change, modification, replacement and modification can be carried out to these embodiments, this
The range of invention is limited by claim and its equivalent replacement, without creative work improvements introduced etc., should all include
Within protection scope of the present invention.
Claims (1)
1. a kind of photocuring 3D printing method of temperature sensitive culture surface material, it is characterised in that include the following steps：
S1 photocuring 3D printing temperature sensing material) is prepared；
S2) photocuring 3D printing temperature sensing material jet printing on substrate surface and is carried out ultraviolet using photocuring 3D printer
Or laser irradiation grafting curing reaction；
S3) make itself and the further graft curing of base material in band type heated at constant temperature channel；
S4) the temperature sensitive culture surface product utilization ultra-pure water of preparation is cleaned, is dried；
The substrate surface refer to surface it is treated containing abundant hydroxyl or carboxyl and can with silane carry out condensation reaction
Glass, ceramics or plastics surface；The photocuring 3D printer refers to Ployjet class ejection-type photocuring 3D printers, spoke
It penetrates cured light source and uses ultraviolet or laser light source, print the layer thickness control of cured temperature sensitive polymer film in 0.01~0.05mm
Between, vertical print speed control is in 0.1~0.3mm/min；
In S4) in step, temperature sensitive culture surface product is using soaking and washing and the processing of the combined type cleaning of ultrasonic cleaning or individually
Using ultrasonic cleaning；
The soaking and washing time is 8~72h；The frequency of oscillation of ultrasonic cleaning is 20~150KHz, and the time is 5~60min；The light
Solidification 3D printing temperature sensing material includes temperature sensitive monomer, grafted monomers, crosslinking agent, chemical initiator, photoinitiator and solvent,
In, the molar ratio of temperature sensitive monomer and grafted monomers is 95:5~85:In 15 ranges, crosslinking agent is temperature sensitive monomer and grafted monomers
The 0.5%~1% of total moles quantity, chemical initiator are the 0.5%~1% of temperature sensitive monomer and grafted monomers total moles quantity,
Photoinitiator is the 0.5%~2% of temperature sensitive monomer and grafted monomers total moles quantity；The temperature sensitive monomer includes N- isopropyls
Acrylamide or N- isopropyl -2- Methacrylamides or n-isopropyl acrylamide and N- isopropyl -2- Methacrylamides
Blend or n-isopropyl acrylamide and N- isopropyl -2- Methacrylamides oligomer, or be less than by molecular weight
2000 n-isopropyl acrylamide mixes the blend of gained with the oligomer of N- isopropyl -2- Methacrylamides；It is described
Grafted monomers include allyltriethoxysilane, allyltrimethoxysilanis, 2- methacrylics triethoxysilane, 2-
Methacrylic trimethoxy silane, propyl acrylate -3- trimethoxy silanes, propyl acrylate -3- triethoxysilanes, first
The one of which or arbitrary two of base propyl acrylate -3- trimethoxy silanes, propyl methacrylate -3- triethoxysilanes
Kind or two or more mixtures；The crosslinking agent includes diacrylate -1,4-butanediol ester, ethylene glycol dimethacrylate
Ester, N, N- methylene bisacrylamides acid imide, bismaleimide amine substance, the crosslinking agent contain two or more insatiable hungers
And double bond；The chemical initiator includes azodiisobutyronitrile, azobisisoheptonitrile or di-t-butyl peroxide, benzoyl peroxide first
Acyl, cumyl peroxide, potassium peroxydisulfate, ammonium persulfate；The photoinitiator uses can directly decompose production under light illumination
Free radical or anion, the benzoin and its derivatives of cation or acetophenones derivative or benzophenone and its derivative
Object；The solvent is polar aprotic solvent, including n,N-Dimethylformamide, n,N-dimethylacetamide, N- methyl pyrroles
The one or more of which of pyrrolidone, tetrahydrofuran, cyclohexanone, the solvent are the hypotoxicities not reacted with hydroxyl and carboxyl
Close object.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610834643.5A CN106378930B (en) | 2016-09-20 | 2016-09-20 | The photocuring 3D printing method of temperature sensitive culture surface material |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610834643.5A CN106378930B (en) | 2016-09-20 | 2016-09-20 | The photocuring 3D printing method of temperature sensitive culture surface material |
Publications (2)
| Publication Number | Publication Date |
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| WO2020156358A1 (en) * | 2019-01-30 | 2020-08-06 | 高雄医学大学 | Slurry for photocuring 3d printing, preparation method therefor, and method of use thereof |
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