CN106236324B - Improve the corneal graft Cascading Methods of tissue engineering artificial corneal diopter - Google Patents
Improve the corneal graft Cascading Methods of tissue engineering artificial corneal diopter Download PDFInfo
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- CN106236324B CN106236324B CN201610563738.8A CN201610563738A CN106236324B CN 106236324 B CN106236324 B CN 106236324B CN 201610563738 A CN201610563738 A CN 201610563738A CN 106236324 B CN106236324 B CN 106236324B
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
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- A—HUMAN NECESSITIES
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- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
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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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- A61L2430/00—Materials or treatment for tissue regeneration
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Abstract
The invention discloses a kind of corneal graft Cascading Methods for improving tissue engineering artificial corneal diopter, are to make tissue engineering artificial corneal with polymeric biomaterial with 3D printing to plant piece, the corneal graft includes front end corneal graft and rear end corneal graft；The front end corneal graft has the size and curvature similar to patient's cornea, is placed in cornea plant bed；The rear end corneal graft has the size and curvature similar to patient's crystalline lens, is placed between the iris of patient and crystalline lens；With reference to the diopter of front end corneal graft, the curvature of antetheca and rear wall in the corneal graft of rear end is adjusted, whole diopter is increased to normal value, corneal graft cascade is realized, solves the technical issues of single tissue engineering artificial corneal diopter is inadequate.
Description
Technical field
The invention belongs to human body implanted medical device technical fields, and in particular to a kind of raising tissue engineering artificial corneal is bent
The corneal graft Cascading Methods of luminosity.
Background technology
Corneal blindness is the second substantially blindness causing disease for being only second to cataract, and vast majority of people can pass through corneal transplantation in patient
It sees light again.Currently, the therapy of corneal blindness mainly includes the transplanting of people's donation operation on cornea, artificial keratoplasty etc..But
It is due to many factors, the cornea source of China various regions eye bank is in great shortage, and many patients can only passively wait for donation.Artificial angle
The research of film has in foreign countries with heterogeneous material such as poly hydroxy ethyl acrylate (PHEMA）, polymethyl methacrylate
(PMMA）, the artificial cornea that is made for raw material research and development of glass, silica gel etc., but there are poor biocompatibilities, and mechanical characteristic is poor, clinical
Test effect is bad, can not form serialization epithelium, it is impossible to autologous patient tissue it is fully integrated the shortcomings of.Patient's rejection
It significantly, can be with serious complication, as being proliferated film and glaucoma after implantation material discharge, infection, entophthamia, formation.De- cell
Pig cornea also by clinic application because being Xenogeneic, however it remains the risk of graft rejection.If it can construct
The problems such as tissue engineering comea of people's cornea will thoroughly solve corneal graft rejection, donor is insufficient is fully equivalent to, is had wide
Application prospect.
The cornea of people is thin, transparent, is by countless arrangements are close, crisscross webbed collagenous fibres are formed, collagen is fine
The latticed array of dimension rule plays the role of diffraction grating, and light scattering is reduced by destroying interference.The central cornea of people
Mean radius of curvature be 7.8mm (6.7~9.4mm), the total diopter of cornea is about 43.25D.Because it is limited by material, technique
System, tissue engineering comea can not also completely bionical people's cornea accurate labyrinth, the in the wrong of tissue engineering comea can be caused in this way
Luminosity is difficult to meet the requirements.
Invention content
The present invention relates to the corneal graft Cascading Methods for improving tissue engineering artificial corneal diopter, solve single tissue work
The technical issues of journey artificial cornea diopter is inadequate.
It is a kind of improve tissue engineering artificial corneal diopter corneal graft Cascading Methods, be with polymeric biomaterial with
3D printing makes tissue engineering artificial corneal and plants piece, and the corneal graft includes front end corneal graft and rear end corneal graft；Institute
Front end corneal graft is stated with the size and curvature similar to patient's cornea, is placed in cornea plant bed；The rear end corneal graft tool
There are the size and curvature similar to patient's crystalline lens, be placed between the iris of patient and crystalline lens；With reference to front end corneal graft
Diopter adjusts the curvature of antetheca and rear wall in the corneal graft of rear end, and whole diopter is increased to normal value, realizes that cornea is planted
Piece cascades.
Further, the preparation method of the front end corneal graft：
1）Prepare cornea three-dimensional rack：Using 3D biometric print machines, printer is placed in for raw material using multiple copolymer
In barrel, according to preset model, the 3 D stereo stent of cornea shape is printed；
2）Functionalized modification is carried out to three-dimensional rack using ECM：De- cell corneal extracellular matrix dry powder is taken to be dissolved in distilled water
In, nano particle suspension is prepared into, using the mode directly coated by the extracellular matrix particle uniform fold in suspension
To cornea three-dimensional rack surface；
3）The structure of remaining cornea micro-assembly robot method：Patient's cornea is taken to carry out micro-assembly robot processing, is prepared into uniform
Precipitation is collected by centrifugation in fine particle, and biological fibrin glue is added in precipitation, aseptically, is injected into and is modified branch
In frame material, front end corneal graft is made.
In order to meet the needs of artificial cornea optical property, mechanical strength and degradation property, the multiple copolymer be by
Various of monomer is polymerized, and the monomer is several in 6-caprolactone, lactide, trimethylene carbonate or collagen
Combination, mass ratio during use is 6-caprolactone：Lactide：Trimethylene carbonate：Collagen=5-90:5-90:3-50:
5-30。
In order to meet the mechanical strength of artificial cornea, the molecular weight of the multiple copolymer is 3000-50000.
In order to improve the biocompatibility of three-dimensional rack, guarantee cell can survive on stent, the three-dimensional rack
Porosity is 90%, and pore diameter is 200-500 μm.
Further, the outer wall radius of curvature of the three-dimensional rack is 7.7mm, and the radius of curvature of inner wall is 6.7mm, wall thickness
It is 200-400 μm, a diameter of 12mm of front end corneal graft.
Further, the rear end corneal graft includes optical section, main part, anchor point fixed part and anchoring in structure
Hole, the optical region has corneal curvature among glasses, and including antetheca and rear wall, the antetheca is convex lens, and rear wall is
Convex lens or concavees lens；The main part is enclosed in around optical section；At least two anchor point fixed parts, for semi-circular shape, thickness
Less than main part, it is symmetrically distributed in around main part；The anchor hole, positioned at the centre position of anchor point fixed part, to penetrate
The circular hole of anchor point fixed part, a diameter of 0.6-1.2mm.
In order to coordinate the advantage of the 3D printing method of forming, according to the performance of the optical section of glasses, main part and anchor point fixed part
The performance requirement of demand and entire glasses is respectively adjusted the composition of raw materials of three parts, the raw material of the optical section
Include the component of following mass parts, 3-8 parts of oxygen permeability material, 5-10 parts of hydrophilic material, 0.01-0.04 parts of initiator, crosslinking
0.1-0.2 parts of agent；The raw material of the main part includes the component of following mass parts, 12-18 parts of oxygen permeability material, hydrophilic material
4-6 parts, 0.01-0.04 parts of initiator, 0.1-0.2 parts of crosslinking agent；The raw material of the anchor point fixed part includes following mass parts
Component, 2-3 parts of oxygen permeability material, 20-30 parts of hydrophilic material, 0.01-0.02 parts of initiator, 0.1-0.15 parts of crosslinking agent.
Further, the preparation method of the rear end corneal graft：
1）Examination of eyes is carried out to patient, acquires optical data, calculates the correction number of degrees of implantable contact lenses；
2）The threedimensional model of implantable contact lenses is established, and diopter adjusts threedimensional model according to needed for optical section
Section carries out layered shaping to model file；
3）The material of optical section, main part or anchor point fixed part is uniformly mixed respectively, is respectively put into and is preheated to 85-90 DEG C
Barrel one, in barrel two or barrel three；It is successively printed under the control of 3D printing software, obtains the implantable stealthy eye of the present invention
Mirror is finely tuned eyeglass lens power, is stored in physiological saline.
In order to be combined with the diopter of front end corneal graft, whole diopter is increased to normal value, the optical section
Antetheca and rear wall all using convex lens, antetheca radius of curvature 12mm, rear wall radius of curvature 10mm, 200-500 μm of thickness.
Compared with prior art, the advantages and positive effects of the present invention are：The present invention provides a kind of raising organizational projects
The Cascading Methods of artificial cornea diopter, including preparing front end corneal graft and preparing rear end corneal graft, two parts use
The good biomaterial with tissue compatibility, and be molded using 3D printing.The front end corneal graft is with the shape of corneal stent
Formula, and it is implanted into autologous patient keratocyte in the bracket, it is placed in the cornea plant bed of patient.The rear end corneal graft is with stealthy eye
The form of mirror, is placed between the iris of patient and crystalline lens.It is analyzed according to ophthalmology image check and computer data, with reference to front end
The diopter of corneal graft adjusts the curvature of antetheca and rear wall in the corneal graft of rear end, whole diopter is increased to normal value.
The present invention solves single tissue engineering artificial corneal diopter using front end corneal graft and the cascade method of rear end corneal graft
The technical issues of inadequate.
Description of the drawings
Fig. 1 is the implantation position schematic diagram of the corneal graft of the present embodiment；
Fig. 2 is that the two-stage corneal graft cascade of the present embodiment improves the schematic diagram of diopter；
Fig. 3 is the structure diagram of the rear end corneal graft of the present embodiment；
It is marked in figure：Front end cornea plant 10, rear end corneal graft 20, optical section 21, main part 22, anchor point fixed part 23,
Anchor hole 24, iris 30, crystalline lens 40, retina 50.
Specific embodiment
Technical scheme of the present invention is described in further detail With reference to embodiment.The present invention is previously mentioned
Ratio, " part ", if without particularly marking, be subject to weight.
It is a kind of improve tissue engineering artificial corneal diopter corneal graft Cascading Methods, be with polymeric biomaterial with
3D printing makes tissue engineering artificial corneal and plants piece, and the corneal graft includes front end cornea and plants 10 and rear end corneal graft 20；
The front end corneal graft has the size and curvature similar to patient's cornea, is placed in cornea plant bed；The rear end corneal graft
With the size and curvature similar to patient's crystalline lens, it is placed between the iris 30 of patient and crystalline lens 40；According to ophthalmology image
It checks and is analyzed with computer data, with reference to the diopter of front end corneal graft, adjust antetheca and rear wall in the corneal graft of rear end
Whole diopter is increased to normal value by curvature, realizes corneal graft cascade.
1st, the preparation method of the front end corneal graft is：
1）Cornea three-dimensional rack is prepared by raw material of multiple copolymer：Using 3D biometric print machines, multiple copolymer is put
In the barrel of printer, primary printing needs raw material 8-12g, and according to preset model, multiple copolymer material is squeezed
Go out on the print platform of sterilizing glass film plates is loaded with, print the 3 D stereo stent of cornea shape.
With reference to the mean parameter of eye cornea structure, the design parameter of the front end corneal graft three-dimensional rack of the present embodiment is such as
Under：The outer wall radius of curvature of three-dimensional rack is 7.7mm, and the radius of curvature of inner wall is 6.7mm, and wall thickness is 200-400 μm, nose angle
Film plants a diameter of 12mm of piece.
A diameter of 200 μm, extruded velocity 0.022-0.033mm/s of print head, print temperature is 120 DEG C -180 DEG C, excellent
150 DEG C, print speed 6mm/s are selected as, printing thickness is 0.01-0.1mm.The technological parameter of above-mentioned 3D printing can influence three-dimensional
The precision of stent, and influence each other between them, such as（1）Print head diameter directly affects the thickness of printout fiber, also
It is related with the particle size of material.（2）Print temperature influences viscosity, feed pressure, forming and hardening speed of material etc., from
And formed precision is influenced indirectly.（3）Print speed is matched with material extruded velocity, material viscosity, material solidification shaping speed,
It is too fast not easily molded or fracture of wire occur, occur very much material stacking slowly.（4）Thickness is printed, is printing precision index in itself, thickness mistake
Greatly, it is desirable that extruded velocity is high, is difficult to realize when material viscosity is excessively high.Thickness is set and material viscosity, print temperature, printing speed
Degree, extruded velocity are related.
It is 90% to print the obtained porosity of cornea three-dimensional rack, and pore diameter is 200-500 μm.The porosity and
Pore diameter has a significant impact to the biocompatibility of stent, and porosity determines the transmission of nutriment and oxygen, pore diameter
Space needed for cell growth is influenced, the two directly determines that cells containing sequences are survived on stent.
The preparation method of the multiple copolymer is：Various of monomer raw material is taken to mix, catalyst is added in, vacuumizes, nitrogen charging
Gas in closed container, is reacted under the conditions of anhydrous and oxygen-free, and temperature is 130-160 DEG C, reaction time 10-24h；
Product is dissolved in dichloromethane or ethyl acetate after the completion of reaction, adding in methanol or petroleum ether precipitates multiple copolymer, mistake
Filter, is dried in vacuo under the conditions of being deposited in 30-50 DEG C, obtains multiple copolymer.
Molecular weight by reaction temperature and reaction time control multiple copolymer is 3000-50000.Point of multiple copolymer
Son amount influences the mechanical property of three-dimensional rack, tensile strength, elasticity modulus and elongation at break including stent, the present embodiment
Middle-molecular-weihydroxyethyl overall target of mechanical property in the range of 3000-50000 is most excellent, meets cornea clinically to power
Learn all demands of intensity；And slow smaller than molecular weight of polymer degradation rate that molecular weight is big.
Several combinations of the monomer in 6-caprolactone, lactide, trimethylene carbonate or collagen use
When mass ratio be 6-caprolactone：Lactide：Trimethylene carbonate：Collagen=5-90:5-90:3-50:5-30.When only
During using two of which or three kinds, more than mass ratio relationship is still met, such as use 6-caprolactone and lactide dibasic monomer
When, the two mass ratio is 5-90:5-90；When for another example using 6-caprolactone, lactide, collagen termonomer, three's mass
Than for 5-90:5-90:5-30.Wherein described lactide can be levorotatory lactide and/or dextrorotation lactide.
The catalyst is stannous octoate, organic guanidine, metallic zinc, tributyltin chloride, ferric acetyl acetonade, zinc lactate, receives
Rice zinc oxide, taurine, ethyl alcohol iron, normal propyl alcohol iron or n-butanol iron.The quality of catalyst is the 0.1%-1% of material quality, excellent
It is selected as 0.5%.
2）Functionalized modification is carried out to three-dimensional rack using ECM：De- cell corneal extracellular matrix dry powder is taken to be dissolved in distilled water
In, particle suspension is prepared into, above-mentioned three-dimensional rack is soaked into suspension, it will be in suspension using the mode directly coated
Extracellular matrix particle uniform fold to cornea three-dimensional rack surface, saved backup at 4 DEG C.
The preparation method of the de- cell corneal extracellular matrix dry powder is：Fresh animal cornea is put in physiological saline
In clean drain after be cut into small pieces, using hydrogen peroxide dipping, the mass concentration of hydrogen peroxide is 3%, soaking time 10-24h；Again
It is cleaned up using distilled water and tri-distilled water, after Bacteria Culture is negative, by fritter cornea using ultra micro wet pulverizing, at 4 DEG C
Under the conditions of centrifuge in three times, centrifugation time is respectively 10-20min, and centrifugal speed is respectively 1000rmp-8000rmp, and three times
The rate of centrifugation is incremental；Precipitation is collected, freeze-drying is prepared into de- cell corneal extracellular matrix nanoscale dry powder, average
A diameter of 130-180nm is preserved at 4 DEG C.
3）The structure of remaining cornea micro-assembly robot method：Patient's cornea is taken, removes degradation degeneration tissue, is retained " relatively just
Often " tissue is spare.Remaining cornea tissue is aseptically transferred in the culture dish in superclean bench, is cut into small pieces,
It is transferred in the vial disinfected, the clostridiopetidase A that 1-5 μ L mass concentrations are 0.1%-1% is added in, for cornea tissue to be detached
It as individual cells, fully shreds, is prepared into uniform remaining cornea tissue suspension, by the strainer filtering of 300 μ m diameters,
Centrifugation discards supernatant collection precipitation, 1-5mL biological fibrin glues is added in precipitation, for adhesion ECM, prevent its exudation；
Under aseptic condition, it is injected into and is modified in timbering material.
The front end corneal graft of the present embodiment uses multipolymer synthetic material, is printed using 3D biometric prints machine
Cornea three-dimensional rack disclosure satisfy that the demand of artificial cornea optical property, mechanical strength and degradation property.Utilize de- cell canthus
Theca cell epimatrix corneal three-dimensional rack carries out functionalized modification so that the canthus membrane support after modification has good extracellular
Matrix microenvironment under the premise of with excellent mechanical performances, obtains more outstanding biocompatibility, and can be further
Regulation and control of the extracellular matrix to cell behavior are played, promote the effect of cell migration, proliferation and differentiation, so that after modification
Stent, which has, preferably promotes the regenerated effect of cornea.Meanwhile the processing of patient remnants' cornea micro-assembly robotization can obtain self
Seed cell, promote " the optimal microenvironment " of tissue engineering comea, have quickly to the application prospect of clinic conversion.
2nd, the preparation method of the rear end corneal graft is：
As shown in figure 3, the rear end corneal graft, includes optical section 21, main part 22, anchor point fixed part 23 in structure
And anchor hole 24.The optical region has corneal curvature, for correction of ophthalmic refractive degree among glasses.The optical section
Including antetheca and rear wall, antetheca is convex lens, and rear wall can be convex lens or concavees lens.The main part is enclosed in optical section week
It encloses, lens surface is attached to after implantation, be used to support optical section.At least two anchor point fixed parts, are semi-circular shape, and thickness is small
It in main part, is symmetrically distributed in around main part, lens surface is closely attached to after implantation, prevent glasses from sliding.It is described
Anchor hole is located at the centre position of anchor point fixed part, to penetrate the circular hole of anchor point fixed part, a diameter of 0.6-1.2mm, for noting
Enter hydrophilic substance, glasses is made to be detached with crystalline lens, if necessary to replace rear end corneal graft, cancelled using anchor hole inverting function
Adherency, facilitates taking-up glasses.
The anchor point fixed part of rear end corneal graft described in the present embodiment is the extension of main part, and size is smaller, and thickness
It is approximately the half of main part thickness less than main part, it is easier to closely attach, can effectively prevent implantable hidden with crystalline lens
Shape glasses slide in eyes.The anchor hole is the micropore for penetrating anchor point fixed part, can be noted by anchor hole to glasses inside
Enter the hydrophilic substances such as hydrogel, glasses can detach rapidly with crystalline lens, facilitate and take out glasses, and therefore, the present embodiment is planted
Enter contact lenses with reverse detachment function.
The main part can be rectangle or circle, and the thickness of main part is 100-800 μm.The radius of anchor point fixed part
It it is 500-2000 μm, thickness is 20-500 μm.
The copolymer material for preparing material and using high oxygen permeability, high-hydrophilic of glasses, can be chronically implanted in the present embodiment,
Good biocompatibility, immunological rejection will not be caused for a long time in vivo by being placed in.The preparation method of glasses uses the 3D printing method of forming.
In the 3D printing method of forming, raw material is different used by the optical sections of glasses, main part and anchor point fixed part, can basis
The performance requirement of three parts and the performance requirement of entire glasses, are respectively adjusted composition of raw materials.Institute in the present embodiment
The raw material for stating optical section includes the component of following mass parts, 3-8 parts of oxygen permeability material, 5-10 parts of hydrophilic material, initiator
0.01-0.04 parts, 0.1-0.2 parts of crosslinking agent；The raw material of the main part includes the component of following mass parts, oxygen permeability material
12-18 parts, 4-6 parts of hydrophilic material, 0.01-0.04 parts of initiator, 0.1-0.2 parts of crosslinking agent；The original of the anchor point fixed part
Material includes the component of following mass parts, and 2-3 parts of oxygen permeability material, 0.01-0.02 parts of initiator, is handed over 20-30 parts of hydrophilic material
Join 0.1-0.15 parts of agent.Feature is to increase hydrophilic component in anchor point fixed part, improves the hydrophily of this part, carries
High and lenticular stickiness；On this basis, oxygen permeability component is increased in main part, improves the oxygen flow of this part
Property, conducive to eye and extraneous gas exchanges.The specific steps are：
1）Examination of eyes is carried out to patient, acquires optical data, the correction number of degrees of computational back-end corneal graft；
2）The threedimensional model of rear end corneal graft is established, and diopter adjusts threedimensional model according to needed for optical section
Section carries out layered shaping to model file；
3）The material of optical section, main part or anchor point fixed part is uniformly mixed respectively, is respectively put into and is preheated to 85-90 DEG C
Barrel one, in barrel two or barrel three；It is successively printed under the control of 3D printing software, the rear end cornea for obtaining the present invention is planted
Piece removes rear end corneal graft from printing substrate, and polishing treatment is finely tuned eyeglass lens power, is stored in physiological saline.
3D printing method can be printed in the different parts of same glasses using different materials, enhance difference
The function at position.And 3D printing method can calculate glasses diopter, be edited into 3D models, beat according to corneal curvature
Print can one-pass molding, eliminate the grinding processing to glasses, precision higher avoids human error, and operation is easier.
Oxygen permeability material described above for methacryloxymethyl trimethyl silane, γ-（Methacryloxypropyl）Propyl
The one or two of trimethoxy silane；The hydrophilic material is hydroxyethyl methacrylate（PHEMA）, N- vinyl pyrroles
Alkanone（PNVP）, acrylic acid etc., polyethylene glycol at least one.The initiator can be azodiisobutyronitrile, peroxidating two
Benzoyl or azobisisoheptonitrile；The crosslinking agent can be N, and N- methylene bisacrylamide acid amide, divinylbenzene or two are different
Cyanate.
Embodiment 1
Front end corneal graft is prepared with following steps：
（1）The synthesis of ternary polymerization material：40% 6-caprolactone monomer, 40% lactide monomer is taken by mass percentage
Collagen monomer with 20% is put into for raw material in consersion unit, and the stannous octoate zinc of material quality 0.5% input by addition is made
It for catalyst, vacuumizes, inflated with nitrogen, in closed polymerization unit, is reacted under the conditions of 160 DEG C for 24 hours, you can obtain molecular weight
For 32838 multiple copolymer.
（2）Print cornea three-dimensional rack：Using above-mentioned ternary polymerization material as raw material, 3D biometric print machines, structure are utilized
The 3-D solid structure body of people's cornea shape, the pore diameter of structure is 350 μm, porosity 90%, and outer wall radius of curvature is
7.7mm, the radius of curvature of inner wall are 6.7mm, and wall thickness is 300 μm, diameter 12mm, obtain can load cells cornea three-dimensional branch
Frame.A diameter of 200 μm, extruded velocity 0.033mm/s of print head, print temperature are 150 DEG C, print speed 6mm/s, printing
Thickness is 0.01mm.
（3）Functionalized modification is carried out to 3D printing stent using de- cell corneal extracellular matrix：Take fresh pig eye canthus
Film, physiological saline is cleaned drain after be cut into small pieces, impregnate 3% hydrogen peroxide 20 hours, after distilled water and tri-distilled water clean up,
By cornea fritter ultra micro wet pulverizing, 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；Precipitation is collected, freeze-drying, which is prepared at 4 DEG C of dry powder, to be preserved, and is done
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 into nanometer
Grain suspension, printed corneal stent is soaked into suspension, and makes the extracellular base in suspension with the mode of coating
Matter particle is uniformly distributed in corneal stent surface, is saved backup at 4 DEG C.
（4）The structure of remaining cornea micro-assembly robot method：If take the remaining cornea after the full excision of corneal injury patient cornea
It is dry, degradation degeneration tissue is removed, it is spare to retain " relatively normal " tissue.Remaining cornea is aseptically transferred to ultra-clean work
Make in platform, be cut into small pieces in culture dish, be then transferred into the vial disinfected, add in the glue that 3 μ L mass concentrations are 1%
Protoenzyme fully shreds, and is prepared into uniform remaining cornea tissue suspension, by the strainer filtering of 300 μ m diameters, centrifugation is abandoned
Fall supernatant collection precipitation, 5mL biological fibrin glues are added in precipitation, aseptically, is injected into and is modified stent
In material.
Front end corneal graft manufactured in the present embodiment, tensile strength 2.38MPa, elasticity modulus 1.6MPa are completely full
Demand of the sufficient organizational project cornea to mechanical strength.
The diopter of front end corneal graft manufactured in the present embodiment：
The refractive index of normal eye's anterior surface of cornea is about 1.376, and the refractive index of the aqueous humor contacted with posterior surface of cornea
It is 1.336.Front refracting power=1000 (1.376-1)/7.7=48.83D, behind refracting power=1000 (1.336-1.376)/
6.7=- 5.97D, therefore, the equivalent refraction power of entire cornea is approximately 43D.
Use proportion 1.008g/cm3Physiological saline simulation aqueous humor, before surveying the present embodiment through RL081630 vertometers
It is 32.35D to hold corneal graft diopter, and about 11D is differed with normal eye's cornea.
Embodiment 2
Rear end corneal graft is prepared with following steps, step is：
1st, examination of eyes is carried out to patient, acquires optical data, the correction number of degrees of computational back-end corneal graft.
The simplified formula of corneal curvature calculating for needing selection is as follows：
（Horizontal curvature+vertical curvature）/ 2*110%=needs the corneal curvature of selection
Such as：Horizontal curvature is 7.8, vertical curvature 7.5, and required eyeglass base curve is： (7.8+7.5)/2*110%=8.4
2nd, 3D printing rear end corneal graft.
1）The threedimensional model of rear end corneal graft is established, the antetheca and rear wall of optical section are all using convex lens, according to patient
Threedimensional model is adjusted glasses diopter in demand, then carries out layered shaping to model file.Rear end corneal graft can
It is designed to structure shown in Fig. 3, three anchor point fixed parts 3 are distributed in circular 2 periphery of main part, anchor point fixed part at equal intervals
3 radius is 1000 μm, can closely be attached to lens surface；
2）Prepared by raw material performed polymer, optical section includes hydroxyethyl methacrylate 5g, n-vinyl pyrrolidone 5g, first
Base alkene pivaloyloxymethyl triethoxysilane 7g, initiator azodiisobutyronitrile 0.03g and crosslinking agent N, N- methylene bisacrylamide
Sour amide 0.15g is uniformly mixed, and is quickly adding into the barrel one for the 3D printing nozzle for being preheated to 90 DEG C；
Main part includes hydroxyethyl methacrylate 2g, n-vinyl pyrrolidone 3g, three second of methyl alkene pivaloyloxymethyl
Oxysilane 16g, initiator azodiisobutyronitrile 0.03g and crosslinking agent N, N- methylene bisacrylamide acid amide 0.15g mixing are equal
It is even, it is quickly adding into the barrel two for the 3D printing nozzle for being preheated to 90 DEG C；
Anchor point fixed part includes hydroxyethyl methacrylate 14g, n-vinyl pyrrolidone 13g, methyl alkene acyloxy first
Ethyl triethoxy silicane alkane 2g, initiator azodiisobutyronitrile 0.03g and crosslinking agent N, N- methylene bisacrylamide acid amide 0.15g are mixed
It closes uniformly, is quickly adding into the barrel three for the 3D printing nozzle for being preheated to 90 DEG C；
3）It is successively printed under the control of 3D printing software, obtains rear end corneal graft.
4) the above-mentioned rear end corneal graft being prepared is processed by shot blasting, finely tunes eyeglass lens power.
3rd, rear end corneal graft is placed in physiological saline and impregnates, preserves, and physiological saline pH value is 7.4.
Calculate the rear end corneal graft diopter of the present embodiment：The antetheca of optical section and rear wall are all using convex in the present embodiment
Lens, antetheca radius of curvature 12mm, rear wall radius of curvature 10mm, 200-500 μm of thickness.Use proportion 1.008g/cm3Physiology
Salt water modeling aqueous humor, the rear end corneal graft diopter that the present embodiment is surveyed through RL081630 vertometers is 11.05D, with reference to preceding
It is 32.35D to hold corneal graft diopter, and therefore, the equivalent refraction power of entire cornea is approximately 43D.
Above example is only several in the several preferred embodiment of the present invention, it is noted that the present invention is not limited to
Above-described embodiment；It for those of ordinary skill in the art, still can be to the technical solution recorded in previous embodiment
It modifies or equivalent replacement is carried out to which part technical characteristic；And these modifications or substitutions, do not make relevant art side
The essence of case is detached from the spirit and scope of claimed technical solution of the invention.
Claims (9)
1. a kind of corneal graft Cascading Methods for improving tissue engineering artificial corneal diopter, which is characterized in that be with macromolecule
Biomaterial makes tissue engineering artificial corneal with 3D printing and plants piece, and the corneal graft includes front end corneal graft and rear end angle
Film plants piece；The front end corneal graft has the size and curvature similar to patient's cornea, can be placed in cornea plant bed；After described
It holds corneal graft that there is the size and curvature similar to patient's crystalline lens, can be placed between the iris of patient and crystalline lens；Knot
The diopter of front end corneal graft is closed, the curvature of antetheca and rear wall in the corneal graft of rear end is adjusted, whole diopter is increased to
Normal value realizes corneal graft cascade；
The preparation method of the front end corneal graft：
1）Prepare cornea three-dimensional rack：Using 3D biometric print machines, the barrel of printer is placed in for raw material using multiple copolymer
In, according to preset model, print the 3 D stereo stent of cornea shape；
2）Functionalized modification is carried out to three-dimensional rack using ECM：De- cell corneal extracellular matrix dry powder is taken to be dissolved in distilled water, is made
It is standby into nano particle suspension, using the mode directly coated by the extracellular matrix particle uniform fold in suspension to cornea
Three-dimensional rack surface；
3）The structure of remaining cornea micro-assembly robot method：Patient's cornea is taken to carry out micro-assembly robot processing, is prepared into uniform subtle
Precipitation is collected by centrifugation in particle, and biological fibrin glue is added in precipitation, aseptically, is injected into and is modified stent material
In material, front end corneal graft is made.
2. corneal graft Cascading Methods according to claim 1, which is characterized in that the multiple copolymer is by a variety of lists
Body is polymerized, several combinations of the monomer in 6-caprolactone, lactide, trimethylene carbonate or collagen,
Mass ratio during use is 6-caprolactone：Lactide：Trimethylene carbonate：Collagen=5-90:5-90:3-50:5-30.
3. corneal graft Cascading Methods according to claim 2, which is characterized in that the molecular weight of the multiple copolymer is
3000-50000。
4. corneal graft Cascading Methods according to claim 1, which is characterized in that the porosity of the three-dimensional rack is
90%, pore diameter is 200-500 μm.
5. corneal graft Cascading Methods according to claim 1, which is characterized in that the outer wall curvature of the three-dimensional rack half
Diameter is 7.7mm, and the radius of curvature of inner wall is 6.7mm, and wall thickness is 200-400 μm, a diameter of 12mm of front end corneal graft.
6. corneal graft Cascading Methods according to claim 1, which is characterized in that the rear end corneal graft is in structure
Including optical section, main part, anchor point fixed part and anchor hole, the optical region has corneal curvature among glasses, including
Antetheca and rear wall, the antetheca are convex lens, and rear wall is convex lens or concavees lens；The main part is enclosed in around optical section；
At least two anchor point fixed parts, are semi-circular shape, and thickness is less than main part, is symmetrically distributed in around main part；The anchoring
Hole, positioned at the centre position of anchor point fixed part, to penetrate the circular hole of anchor point fixed part, a diameter of 0.6-1.2mm.
7. corneal graft Cascading Methods according to claim 6, which is characterized in that the raw material of the optical section includes as follows
The component of mass parts, 3-8 parts of oxygen permeability material, 5-10 parts of hydrophilic material, 0.01-0.04 parts of initiator, crosslinking agent 0.1-0.2
Part；The raw material of the main part includes the component of following mass parts, and 12-18 parts of oxygen permeability material, draws 4-6 parts of hydrophilic material
Send out 0.01-0.04 parts of agent, 0.1-0.2 parts of crosslinking agent；The raw material of the anchor point fixed part includes the component of following mass parts, oxygen flow
2-3 parts of material of property, 20-30 parts of hydrophilic material, 0.01-0.02 parts of initiator, 0.1-0.15 parts of crosslinking agent.
8. corneal graft Cascading Methods according to claim 6, which is characterized in that the preparation side of the rear end corneal graft
Method：
1）Examination of eyes is carried out to patient, acquires optical data, calculates the correction number of degrees of implantable contact lenses；
2）The threedimensional model of implantable contact lenses is established, and threedimensional model is adjusted in diopter according to needed for optical section,
Layered shaping is carried out to model file；
3）The material of optical section, main part or anchor point fixed part is uniformly mixed respectively, is respectively put into and is preheated to 85-90 DEG C of material
In cylinder one, barrel two or barrel three；It is successively printed under the control of 3D printing software, obtains implantable contact lenses, finely tune glasses
The number of degrees are stored in physiological saline.
9. corneal graft Cascading Methods according to claim 6, which is characterized in that the antetheca and rear wall of the optical section are all
Using convex lens, antetheca radius of curvature 12mm, rear wall radius of curvature 10mm, 200-500 μm of thickness.
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| EP3781083A1 (en) * | 2018-04-17 | 2021-02-24 | Alcon Inc. | Making an implant for an eye |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4892543A (en) * | 1989-02-02 | 1990-01-09 | Turley Dana F | Intraocular lens providing accomodation |
| EP1457171A2 (en) * | 1993-08-27 | 2004-09-15 | Medevec Licensing B.V. | Accommodating intraocular lens |
| CN101172070A (en) * | 2006-10-30 | 2008-05-07 | 卡尔豪恩视觉公司 | Piggyback lenses |
| 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 |
-
2016
- 2016-07-18 CN CN201610563738.8A patent/CN106236324B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4892543A (en) * | 1989-02-02 | 1990-01-09 | Turley Dana F | Intraocular lens providing accomodation |
| EP1457171A2 (en) * | 1993-08-27 | 2004-09-15 | Medevec Licensing B.V. | Accommodating intraocular lens |
| CN101172070A (en) * | 2006-10-30 | 2008-05-07 | 卡尔豪恩视觉公司 | Piggyback lenses |
| 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 |
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