CN106139249A - A kind of preparation method of multipolymer artificial cornea - Google Patents
A kind of preparation method of multipolymer artificial cornea Download PDFInfo
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- CN106139249A CN106139249A CN201610563733.5A CN201610563733A CN106139249A CN 106139249 A CN106139249 A CN 106139249A CN 201610563733 A CN201610563733 A CN 201610563733A CN 106139249 A CN106139249 A CN 106139249A
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- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3604—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix characterised by the human or animal origin of the biological material, e.g. hair, fascia, fish scales, silk, shellac, pericardium, pleura, renal tissue, amniotic membrane, parenchymal tissue, fetal tissue, muscle tissue, fat tissue, enamel
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Abstract
The invention discloses the preparation method of a kind of multipolymer artificial cornea, step includes: 1) synthesis multi-component copolymer material: takes the mixing of various of monomer raw material, adds catalyst, evacuation, inflated with nitrogen, react, prepare multiple copolymer in airtight container；2) cornea three-dimensional rack is prepared: using 3D biometric print machine, use multiple copolymer is raw material, prints the 3 D stereo support of cornea shape；3) utilize ECM that three-dimensional rack carries out functionalized modification: to take de-cell corneal extracellular matrix dry powder and be dissolved in distilled water, by the extracellular matrix granule uniform fold in suspension to cornea three-dimensional rack surface；4) structure of remaining cornea micro-assembly robot method: take patient's cornea and carry out micro-assembly robot process, be injected into and be modified in timbering material, prepares a kind of excellent in mechanical performance, the artificial cornea that biocompatibility is good.
Description
Technical field
The invention belongs to human body implanted medical device technology technical field, particularly relate to a kind of multiple aggregation utilizing synthesis
Thing prepares the method for artificial cornea.
Background technology
People's corneal stroma accounts for the 90% of corneal thickness, mainly by the extracellular matrix such as keratocyte and collagen fiber
Composition is regularly arranged to be formed, transparent and have certain radius of curvature, beneficially light by and warpage, maintain whole cornea
Transparency and the aspect such as bent light rate there is vital effect.The sick blind patient of most corneal stromas all can pass through angle
Film transplant operation is cured, but due to contribute the height scarcity of cornea cause Most patients to can not get donor's cornea and cannot
Recover lost eyesight, cause many cornea Diseases to be chronically at and wait that donor is contributed and in blind misery.Therefore exploitation one is come
Source is convenient, and alternative donor is contributed the artificial cornea of cornea and just become increasingly important and urgent.
Cornea,artificial's timbering material mainly by synthesising biological material and to ECM(extracellular matrix) relevant biomaterial group
Become.The advantage of synthesising biological material is to provide the biomechanical property of excellence, and endless supply, transmittance is high and plastic
Property strong feature, shortcoming is that hydrophilic is poor, lacks biological activity, and may induce the problems such as a series of inflammatory reaction.Close
Become and biomaterial add ECM composition, it is possible to for seed cell create one contribute to cell adhesion, breed or break up natural
Microenvironment.
3D printing technique is a rapid shaping technique based on ejection-type, can be easily controlled local material composition,
Microstructure and surface characteristic, may apply in the preparation of tissue engineering comea.
Summary of the invention
The present invention relates to the preparation method of a kind of multipolymer artificial cornea, obtain a kind of excellent in mechanical performance, biological
The artificial cornea that the compatibility is good.
The technical scheme used for achieving the above object is: the preparation method of a kind of multipolymer artificial cornea, step
Including:
1) synthesis multi-component copolymer material: taking the mixing of various of monomer raw material, add catalyst, evacuation, inflated with nitrogen, in airtight appearance
Device reacts, prepares multiple copolymer；
2) cornea three-dimensional rack is prepared: use 3D biometric print machine, be the material that raw material is placed in printer by above-mentioned multiple copolymer
In Tong, according to model set in advance, print the 3 D stereo support of cornea shape；
3) utilize ECM that three-dimensional rack carries out functionalized modification: to take de-cell corneal extracellular matrix dry powder and be dissolved in distilled water, system
Standby one-tenth nano-particle suspension, uses the mode directly coated by the extracellular matrix granule uniform fold in suspension to cornea
Three-dimensional rack surface；
4) structure of remaining cornea micro-assembly robot method: take patient's cornea and carry out micro-assembly robot process, be prepared as the trickleest
Granule, centrifugal collecting precipitation, precipitation adds biological fibrin glue, aseptically, is injected into and is modified support material
In material, prepare artificial cornea.
In order to control the molecular weight of multiple copolymer, in described step 1), reaction temperature is 130-160 DEG C, and the response time is
10-24h。
In order to meet the mechanical strength of artificial cornea, in described step 1), the molecular weight of multiple copolymer is 3000-
50000。
In order to meet the demand of artificial cornea's optical property, mechanical strength and degradation property, monomer choosing in described step 1)
Several combinations in 6-caprolactone, lactide, trimethylene carbonate or ethylene glycol, mass ratio during use is, ε-own interior
Ester: lactide: trimethylene carbonate: ethylene glycol=5-90:5-90:3-50:5-30.
In order to accelerate multiple aggregation reaction, in described step 1), catalyst is stannous octoate, organic guanidine, metallic zinc, three fourths
Base stannic chloride, ferric acetyl acetonade, zinc lactate, nano zine oxide, taurine, ethanol ferrum, normal propyl alcohol ferrum or n-butyl alcohol ferrum；Catalyst
The 0.1%-1% that quality is raw materials quality.
Further, described step 2) in a diameter of 200 μm of printhead, extruded velocity is 0.022-0.033mm/s, prints
Temperature is 120 DEG C-180 DEG C, and print speed is 6mm/s, and printing thickness is 0.01-0.1mm.
In order to improve the biocompatibility of three-dimensional rack, it is ensured that cell can be survived on support, described step 2) in beat
The porosity of the cornea three-dimensional rack that print obtains is 90%, and pore diameter is 200-500 m.
Further, in described step 3), the preparation method of de-cell corneal extracellular matrix dry powder is: by fresh animal
Cornea be put in normal saline clean drain after be cut into small pieces, use hydrogen peroxide dipping；Re-use distilled water and tri-distilled water is clear
Wash clean, after antibacterial culturing is negative, uses fritter cornea ultra micro waterproof pulverization, centrifugation, collects precipitation, lyophilization
It is prepared as de-cell corneal extracellular matrix nanoscale dry powder, preserves at 4 DEG C.
In order to improve centrifugal effect, reducing the particle diameter of de-cell corneal extracellular matrix granule, described being centrifuged is at 4 DEG C of bars
Being centrifuged in three times under part, centrifugation time is respectively 10-20min, and centrifugal speed is respectively 1000rmp-8000rmp, and three times from
The speed of the heart is incremented by.
Compared with prior art, advantages of the present invention and good effect are: the method comprises the steps of firstly, preparing one and have excellent power
Learn the multipolymer material of performance, utilize 3D printing technique to prepare cornea three-dimensional rack with multipolymer for raw material；Then
Utilize de-cell corneal extracellular matrix dry powder (ECM) that three-dimensional rack carries out functionalized modification, use 3D printing type or use
Directly coating mode by extracellular matrix granule uniform fold to cornea three-dimensional rack surface；Finally by the remaining cornea of patient
Carry out " micro-assembly robotization process ", also will be compound in cornea three-dimensional rack by auto corneal " seed cell ", promote artificial angle
The optimum microenvironment of film, it is to avoid the In vitro culture link of seed cell, has the application prospect that rapid clinical converts.
Detailed description of the invention
Below in conjunction with detailed description of the invention, technical scheme is described in further detail.The present invention is previously mentioned
Ratio, " part ", without special labelling, be all as the criterion with weight.
The invention provides the preparation method of a kind of artificial cornea, the method utilizing chemosynthesis, a kind of mechanical property of preparation
The adjustable synthesising biological material of energy, utilizes 3D printing technique to print artificial cornea's support, in order to improve its biocompatibility, uses
ECM carries out biological function sex modification so that it is on the premise of having excellent mechanical performances, obtains the biological activity of excellence.Step
Including:
1) synthesis multi-component copolymer material: taking the mixing of various of monomer raw material, add catalyst, evacuation, inflated with nitrogen, in airtight appearance
In device, reacting under the conditions of anhydrous and oxygen-free, temperature is 130-160 DEG C, and the response time is 10-24h；To produce after having reacted
Thing is dissolved in dichloromethane or ethyl acetate, adds methanol or petroleum ether makes multiple copolymer precipitate, filter, will be deposited in 30-
It is vacuum dried under the conditions of 50 DEG C, obtains multiple copolymer.
The molecular weight being controlled multiple copolymer by reaction temperature and response time is 3000-50000.Dividing of multiple copolymer
Son amount affects the mechanical property of three-dimensional rack, including hot strength, elastic modelling quantity and the elongation at break of support, the present embodiment
Middle-molecular-weihydroxyethyl aggregative indicator of mechanical property in the range of 3000-50000 is the most excellent, meets cornea clinically to power
Learn all demands of intensity；And slow less than molecular weight of the big polymer degradation rate of molecular weight.
It is several that monomer described in the present embodiment is selected from 6-caprolactone, lactide, trimethylene carbonate or ethylene glycol
Combination, mass ratio during use is, 6-caprolactone: lactide: trimethylene carbonate: ethylene glycol=5-90:5-90:3-50:5-
30.When only using two of which or when three kinds, still meet above mass ratio relation, such as, use 6-caprolactone and lactide two
During unit's monomer, the two mass ratio is 5-90:5-90；And for example use 6-caprolactone, lactide, trimethylene carbonate termonomer
Time, three's mass ratio is 5-90:5-90:3-50.Wherein said lactide can be levorotatory lactide and/or dextrorotation lactide.
Described catalyst is stannous octoate, organic guanidine, metallic zinc, tributyltin chloride, ferric acetyl acetonade, zinc lactate, receives
Rice zinc oxide, taurine, ethanol ferrum, normal propyl alcohol ferrum or n-butyl alcohol ferrum.The quality of catalyst is the 0.1%-1% of raw materials quality, excellent
Elect 0.5% as.
2) cornea three-dimensional rack is prepared: use the 3D biometric print machine of stepping motorized motions, by above-mentioned multiple copolymer be
Raw material is placed in the barrel of printer, and once printing needs raw material 8-12g, according to model set in advance, by multiple copolymer
Material is extruded on the print platform being loaded with sterilizing glass film plates, prints the 3 D stereo support of cornea shape.Printhead is straight
Footpath is 200 μm, and extruded velocity is 0.022-0.033mm/s, and print temperature is 120 DEG C-180 DEG C, preferably 150 DEG C, prints speed
Degree is 6mm/s, and printing thickness is 0.01-0.1mm.
The technological parameter that above-mentioned 3D prints can affect the precision of three-dimensional rack, and influences each other between them, and such as (1) is beaten
Print head diameter, directly affects the thickness of printout fiber, also relevant with the size of material.(2) print temperature, has influence on
The viscosity of material, feed pressure, forming and hardening speed etc., thus remote-effects formed precision.(3) print speed, squeezes with material
Go out speed, material viscosity, material curing molding speeds match, too fast not easy-formation or fracture of wire occurs, material stacking occurs the most slowly.
(4) printing thickness, itself be printing precision index, thickness is excessive, it is desirable to extruded velocity is high, is difficult to reality when material viscosity is too high
Existing.Thickness arranges relevant to material viscosity, print temperature, print speed, extruded velocity.
Printing the porosity of cornea three-dimensional rack obtained is 90%, and pore diameter is 200-500 m, described porosity and
The biocompatibility of support is had a significant impact by pore diameter, and porosity determines nutrient substance and the transmission of oxygen, pore diameter
Affecting cell growth requisite space, the two directly determines that cells containing sequences is survived on support.
3) utilize ECM that three-dimensional rack carries out functionalized modification: to take de-cell corneal extracellular matrix dry powder and be dissolved in distilled water
In, it is prepared as nano-particle suspension, above-mentioned three-dimensional rack is soaked in suspension, use the mode of directly coating by suspendible
Extracellular matrix granule uniform fold in liquid, to cornea three-dimensional rack surface, saves backup at 4 DEG C.
The preparation method of described de-cell corneal extracellular matrix dry powder is: fresh animal cornea is put in normal saline
In clean drain after be cut into small pieces, use hydrogen peroxide dipping, the mass concentration of hydrogen peroxide is 3%, and soak time is 10-24h；Again
Use distilled water and tri-distilled water to clean up, after antibacterial culturing is negative, fritter cornea is used ultra micro waterproof pulverization, at 4 DEG C
Under the conditions of be centrifuged in three times, centrifugation time is respectively 10-20min, and centrifugal speed is respectively 1000rmp-8000rmp, and three times
Centrifugal speed is incremented by；Collecting precipitation, lyophilization is prepared as de-cell corneal extracellular matrix nanoscale dry powder, averagely
A diameter of 130-180nm, preserves at 4 DEG C.
4) structure of remaining cornea micro-assembly robot method: take patient's cornea, remove degradation degeneration tissue, retain and " align mutually
Often " tissue is standby.Remnants cornea tissue is carried out micro-assembly robot process, aseptically transfers to the training in superclean bench
Supporting in ware, be cut into small pieces, transfer in the vial disinfected, adding 1-5 L mass concentration is the collagenase of 0.1%-1%, uses
In cornea tissue is separated into individual cells, fully shred, be prepared as uniform remaining cornea tissue suspension, by 300
The strainer filtering of m diameter, centrifugal, discard supernatant collection precipitation, precipitation adds 1-5mL biological fibrin glue, for adhesion
ECM, prevents it from oozing out；Aseptically, it is injected into and is modified in timbering material.
The present invention use multipolymer synthetic material disclosure satisfy that artificial cornea's optical property, mechanical strength and
The demand of degradation property.Utilizing 3D biometric print machine to print cornea three-dimensional rack, precision is high.Utilize de-cell cornea extracellular
Substrate corneal three-dimensional rack carries out functionalized modification so that the cornea support after modification has good extracellular matrix micro-loop
Border, on the premise of having excellent mechanical performances, it is thus achieved that the most outstanding biocompatibility, and can play cell further
The epimatrix regulation and control to cell behavior, the effect promote cell migration, breeding and breaking up, so that the support after Xiu Shiing has
Preferably promote the effect of cornea regeneration.Meanwhile, patient remnants' cornea micro-assembly robotization processes and is obtained in that autologous kind is careful
Born of the same parents, promote " the optimum microenvironment " of tissue engineering comea, have the application prospect quickly converted to clinic.
Embodiment 1
Artificial cornea is prepared with following steps:
(1) synthesis of binary copolymerization material: take the 6-caprolactone monomer of 35% by mass percentage, the lactide monomer of 65% is former
Material puts in consersion unit, and the stannous octoate of addition raw materials quality 0.5% is as catalyst, evacuation, and inflated with nitrogen, airtight
In polymerization unit, under the conditions of 150 DEG C, react 20h, the multiple copolymer that molecular weight is 22938 can be obtained.
(2) print cornea three-dimensional rack: with above-mentioned ternary polymerization material as raw material, utilize biometric print machine, build people
The 3 D stereo structure of cornea shape, the pore diameter of structure is 350 m, and porosity is 90%, and obtaining can the cornea of load cells
Three-dimensional rack.A diameter of 200 μm of printhead, extruded velocity is 0.022mm/s, and print temperature is 120 DEG C, and print speed is 6mm/
S, printing thickness is 0.1mm.
(3) utilize de-cell corneal extracellular matrix that 3D print carriage carries out functionalized modification: to take fresh Oculus sus domestica canthus
Film, normal saline is cleaned after draining and is cut into small pieces, soaks the hydrogen peroxide 20 hours of 3%, after distilled water and tri-distilled water clean up,
By cornea fritter ultra micro waterproof pulverization, being centrifuged in three times under the conditions of 4 DEG C, centrifugation time is followed successively by 10min, 15min, 20min, from
Heart speed is followed successively by 1000rmp, 5000 rmp, 8000rmp；Collecting precipitation, lyophilization is prepared as at 4 DEG C of dry powder preserving, dry
The a diameter of 150-180nm of powder.De-cell corneal extracellular matrix dry powder is dissolved in distilled water, is prepared as nano-particle suspendible
Liquid, is soaked into printed CF in suspension, and by the mode of coating by the extracellular matrix granule in suspension
It is uniformly distributed in CF surface, saves backup at 4 DEG C.
(4) structure of remaining cornea micro-assembly robot method: if taking the remaining cornea after corneal injury patient's cornea excises entirely
Dry, remove degradation degeneration tissue, retain " the most normal " tissue standby.Aseptically transfer to surpass by remnants cornea tissue
In clean workbench, being cut into small pieces, be then transferred in the vial disinfected in culture dish, adding 2 L mass concentrations is
The collagenase of 0.1%, fully shreds, and is prepared as uniform remaining cornea tissue suspension, by the strainer filtering of 300 m diameters,
Centrifugal, discard supernatant collection precipitation, precipitation adds 3mL biological fibrin glue, aseptically, is injected into and is repaiied
In decorations timbering material.
Artificial cornea prepared by the present embodiment, hot strength is 2.3MPa, and elastic modelling quantity is 4.2MPa, fully meets tissue
The engineering cornea demand to mechanical strength.
Embodiment 2
Artificial cornea is prepared with following steps:
(1) synthesis of ternary polymerization material: take the 6-caprolactone monomer of 5%-90%, the lactide of 5%-90% by mass percentage
The trimethylene carbonate monomer of monomer and 3%-50% is that raw material puts in consersion unit, and adding quality is input raw materials quality
0.5% tributyltin chloride as catalyst, evacuation, inflated with nitrogen, in airtight polymerization unit, under the conditions of 158 DEG C
React 20 hours, the multiple copolymer that molecular weight is 48338 can be obtained.
(2) print cornea three-dimensional rack: with above-mentioned ternary polymerization material as raw material, utilize biometric print machine, build people
The 3 D stereo structure of cornea shape, the pore diameter of structure is 250 m, and porosity is 90%, and obtaining can the cornea of load cells
Three-dimensional rack.A diameter of 200 μm of printhead, extruded velocity is 0.028mm/s, and print temperature is 150 DEG C, and print speed is 6mm/
S, printing thickness is 0.05mm.
(3) utilize de-cell corneal extracellular matrix that 3D print carriage carries out functionalized modification: to take fresh Oculus sus domestica canthus
Film, normal saline is cleaned after draining and is cut into small pieces, soaks 3% hydrogen peroxide 18h, after distilled water and tri-distilled water clean up, by cornea
Fritter ultra micro waterproof pulverization, is centrifuged under the conditions of 4 DEG C in three times, and centrifugation time is followed successively by 10min, 10min, 10min, centrifugal speed
It is followed successively by 3000rmp, 6000 rmp, 9000rmp；Collecting precipitation, lyophilization is prepared as at 4 DEG C of dry powder preserving, dry powder diameter
For 130-150nm.De-cell corneal extracellular matrix dry powder is dissolved in distilled water, is prepared as nano-particle suspension, will beat
Printed CF is soaked in suspension, makes the extracellular matrix granule in suspension be uniformly distributed in by the mode of coating
CF surface, saves backup at 4 DEG C.
(4) structure of remaining cornea micro-assembly robot method: if taking the remaining cornea after corneal injury patient's cornea excises entirely
Dry, remove degradation degeneration tissue, retain " the most normal " tissue standby.Aseptically transfer to surpass by remnants cornea tissue
In clean workbench, being cut into small pieces, be then transferred in the vial disinfected in culture dish, adding 3 L mass concentrations is
The collagenase of 0.5%, fully shreds, and is prepared as uniform remaining cornea tissue suspension, by the strainer filtering of 300 m diameters,
Centrifugal, discard supernatant collection precipitation, precipitation adds 4mL biological fibrin glue, aseptically, is injected into and is repaiied
In decorations timbering material.
Artificial cornea prepared by the present embodiment, hot strength is 2.68MPa, and elastic modelling quantity is 3.2MPa, fully meets group
Weaver's journey cornea demand to mechanical strength.
Embodiment 3
Artificial cornea is prepared with following steps:
(1) synthesis of ternary polymerization material: take by mass percentage 40% 6-caprolactone monomer, the lactide monomer of 40% and
The ethylene glycol monomers of 20% is that raw material puts in consersion unit, and input by addition, the zinc lactate of raw materials quality 0.5% is as catalyst,
Evacuation, inflated with nitrogen, in airtight polymerization unit, under the conditions of 160 DEG C, react 24h, can obtain molecular weight is 32838
Multiple copolymer.
(2) print cornea three-dimensional rack: with above-mentioned ternary polymerization material as raw material, utilize 3D biometric print machine, build
The 3 D stereo structure of people's cornea shape, the pore diameter of structure is 350 m, and porosity is 90%, and obtaining can the angle of load cells
Film three-dimensional rack.A diameter of 200 μm of printhead, extruded velocity is 0.033mm/s, and print temperature is 150 DEG C, and print speed is
6mm/s, printing thickness is 0.01mm.
(3) utilize de-cell corneal extracellular matrix that 3D print carriage carries out functionalized modification: to take fresh Oculus sus domestica canthus
Film, normal saline is cleaned after draining and is cut into small pieces, soaks the hydrogen peroxide 20 hours of 3%, after distilled water and tri-distilled water clean up,
By cornea fritter ultra micro waterproof pulverization, being centrifuged in three times under the conditions of 4 DEG C, centrifugation time is followed successively by 15min, 15min, 20min, from
Heart speed is followed successively by 4000rmp, 5000 rmp, 6000rmp；Collecting precipitation, lyophilization is prepared as at 4 DEG C of dry powder preserving, dry
The a diameter of 140-160nm of powder.A certain amount of de-cell corneal extracellular matrix dry powder is dissolved in distilled water, is prepared as nanometer
Grain suspension, is soaked into printed CF in suspension, and makes the extracellular base in suspension by the mode of coating
Matter granule is uniformly distributed in CF surface, saves backup at 4 DEG C.
(4) structure of remaining cornea micro-assembly robot method: if taking the remaining cornea after corneal injury patient's cornea excises entirely
Dry, remove degradation degeneration tissue, retain " the most normal " tissue standby.Remnants cornea is aseptically transferred to ultra-clean work
In station, being cut into small pieces in culture dish, be then transferred in the vial disinfected, adding 5 L mass concentrations is the glue of 1%
Protoenzyme, fully shreds, and is prepared as uniform remaining cornea tissue suspension, by the strainer filtering of 300 m diameters, centrifugal, abandons
Fall supernatant collection precipitation, precipitation adds 5mL biological fibrin glue, aseptically, is injected into and is modified support
In material.
Artificial cornea prepared by the present embodiment, hot strength is 2.38MPa, and elastic modelling quantity is 1.6MPa, fully meets group
Weaver's journey cornea demand to mechanical strength.
Above example is only several in the several preferred implementation of the present invention, it is noted that the invention is not restricted to
Above-described embodiment；For the person of ordinary skill of the art, still can be to the technical scheme described in previous embodiment
Modify, or wherein portion of techniques feature is carried out equivalent；And these amendments or replacement, do not make relevant art side
The essence of case departs from the spirit and scope of claimed technical solution of the invention.
Claims (9)
1. the preparation method of a multipolymer artificial cornea, it is characterised in that step includes:
1) synthesis multi-component copolymer material: taking the mixing of various of monomer raw material, add catalyst, evacuation, inflated with nitrogen, in airtight appearance
Device reacts, prepares multiple copolymer；
2) cornea three-dimensional rack is prepared: use 3D biometric print machine, be the material that raw material is placed in printer by above-mentioned multiple copolymer
In Tong, according to model set in advance, print the 3 D stereo support of cornea shape；
3) utilize ECM that three-dimensional rack carries out functionalized modification: to take de-cell corneal extracellular matrix dry powder and be dissolved in distilled water, system
Standby one-tenth nano-particle suspension, uses the mode directly coated by the extracellular matrix granule uniform fold in suspension to cornea
Three-dimensional rack surface；
4) structure of remaining cornea micro-assembly robot method: take patient's cornea and carry out micro-assembly robot process, be prepared as the trickleest
Granule, centrifugal collecting precipitation, precipitation adds biological fibrin glue, aseptically, is injected into and is modified support material
In material, prepare artificial cornea.
The preparation method of a kind of multipolymer artificial cornea the most according to claim 1, it is characterised in that described step
1) in, reaction temperature is 130-160 DEG C, and the response time is 10-24h.
The preparation method of a kind of multipolymer artificial cornea the most according to claim 1, it is characterised in that described step
1) in, the molecular weight of multiple copolymer is 3000-50000.
The preparation method of a kind of multipolymer artificial cornea the most according to claim 1, it is characterised in that described step
1) monomer several combinations in 6-caprolactone, lactide, trimethylene carbonate or ethylene glycol, mass ratio during use in
For, 6-caprolactone: lactide: trimethylene carbonate: ethylene glycol=5-90:5-90:3-50:5-30.
The preparation method of a kind of multipolymer artificial cornea the most according to claim 1, it is characterised in that described step
1) in catalyst be stannous octoate, organic guanidine, metallic zinc, tributyltin chloride, ferric acetyl acetonade, zinc lactate, nano zine oxide,
Taurine, ethanol ferrum, normal propyl alcohol ferrum or n-butyl alcohol ferrum；The quality of catalyst is the 0.1%-1% of raw materials quality.
6. according to the preparation method of a kind of multipolymer artificial cornea described in any one of claim 1-5, it is characterised in that
Described step 2) in a diameter of 200 μm of printhead, extruded velocity is 0.022-0.033mm/s, and print temperature is 120 DEG C-180
DEG C, print speed is 6mm/s, and printing thickness is 0.01-0.1mm.
The preparation method of a kind of multipolymer artificial cornea the most according to claim 6, it is characterised in that described step
2) porosity printing the cornea three-dimensional rack obtained in is 90%, and pore diameter is 200-500 m.
The preparation method of a kind of multipolymer artificial cornea the most according to claim 6, it is characterised in that described step
3) in, the preparation method of de-cell corneal extracellular matrix dry powder is: is put in normal saline by fresh animal cornea and cleans drip
It is cut into small pieces after Gan, uses hydrogen peroxide dipping；Re-use distilled water and tri-distilled water cleans up, after antibacterial culturing is negative, will
Fritter cornea uses ultra micro waterproof pulverization, centrifugation, collects precipitation, and lyophilization is prepared as de-cell corneal extracellular matrix
Nanoscale dry powder, preserves at 4 DEG C.
The preparation method of a kind of multipolymer artificial cornea the most according to claim 8, it is characterised in that described centrifugal
Being to be centrifuged in three times under the conditions of 4 DEG C, centrifugation time is respectively 10-20min, and centrifugal speed is respectively 1000rmp-8000rmp,
And three centrifugal speed is incremented by.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1460715A (en) * | 2003-05-22 | 2003-12-10 | 上海第二医科大学附属第九人民医院 | Production method of transplanting material |
| CN104546222A (en) * | 2015-02-02 | 2015-04-29 | 深圳华明生物科技有限公司 | Artificial cornea and manufacturing method thereof |
| CN104768586A (en) * | 2012-09-04 | 2015-07-08 | 人类起源公司 | Methods of tissue generation |
| CN104822342A (en) * | 2012-10-08 | 2015-08-05 | 矩阵心血管疾病有限公司 | Method and system for treating biological tissue |
| CN105034369A (en) * | 2015-07-10 | 2015-11-11 | 青岛尤尼科技有限公司 | Three-dimensional (3D) cornea stroma support material and method for constructing three-dimensional cornea stroma support |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1460715A (en) * | 2003-05-22 | 2003-12-10 | 上海第二医科大学附属第九人民医院 | Production method of transplanting material |
| CN104768586A (en) * | 2012-09-04 | 2015-07-08 | 人类起源公司 | Methods of tissue generation |
| CN104822342A (en) * | 2012-10-08 | 2015-08-05 | 矩阵心血管疾病有限公司 | Method and system for treating biological tissue |
| CN104546222A (en) * | 2015-02-02 | 2015-04-29 | 深圳华明生物科技有限公司 | Artificial cornea and manufacturing method thereof |
| CN105034369A (en) * | 2015-07-10 | 2015-11-11 | 青岛尤尼科技有限公司 | Three-dimensional (3D) cornea stroma support material and method for constructing three-dimensional cornea stroma support |
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