CN101380486B - Active regeneration artificial cornea graft and preparation method thereof - Google Patents
Active regeneration artificial cornea graft and preparation method thereof Download PDFInfo
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- CN101380486B CN101380486B CN2008102186940A CN200810218694A CN101380486B CN 101380486 B CN101380486 B CN 101380486B CN 2008102186940 A CN2008102186940 A CN 2008102186940A CN 200810218694 A CN200810218694 A CN 200810218694A CN 101380486 B CN101380486 B CN 101380486B
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Abstract
The invention belongs to the technical field of artificial cornea production and more particularly provides an active regenerating artificial cornea implant and a preparation method thereof. The artificial cornea implant comprises 5-30 shares of chitosan, 5-30 shares of collagen, 0.1-0.5 share of chondroitin-sulfuric acid, 5-10 shares of polyethylene glycol, 5-25 shares of polyvinyl alcohol and 15-80 shares of triple-distilled water; preferably the artificial cornea implant also comprises appropriate amount of cornea cell growing factor, fibronectin and laminin. The preparation method of the active regenerating artificial cornea implant is as below: dissolving the chitosan, the collagen and the chondroitin-sulfuric acid in hydrochloric acid, acetic acid and water respectively, and then infusing the mixture of the components which are agitated uniformly at the room temperature into a mould for drying, and then obtaining the product after water immersion, cleaning and drying. The artificial cornea implant has good biocompatibility, high substance rigidity and good transparency; the preparation method is easy to be operated, the raw materials are easy to be obtained and are cheap, and large-scale industrialized production can be performed.
Description
Technical field
The present invention relates to the technical field that the artificial cornea makes, be specifically related to a kind of active regeneration artificial cornea graft and preparation method thereof.
Background technology
The whole world has 1,000 ten thousand corneal blindness needs of patients to carry out corneal transplantation approximately, China 3,000,000 people that have an appointment.But, only about 3000 examples of allogeneic keratoplasty that China can carry out every year.This mainly is that to carry out the corneal transplantation price more expensive because the allogeneic cornea is a donor, is difficult to obtain donor.The shortage of donor makes development become the technical problem that presses for solution as the artificial cornea of allogeneic cornea succedaneum.
Present artificial cornea is main with nonactive cornea, and this type of cornea is generally by synthetic material such as methyl methacrylate manufacturings such as (MMA), though the implantation of this type of material can make people's vision recover to some extent; But field range is minimum; Be difficult to merge, foreign body sensation is arranged after the implantation, occur corneal solution, leakage of aqueous humor easily, deviate from and complication such as endophthalmitis with enclosing near the eyes to organize mutually; Influence growing into of corneal nerve, be easy to cause xerophthalmia.
Aspect the improving of artificial cornea, tissue engineering comea has appearred in recent years.This type cornea is under the condition of external reinforcement, on artificial cornea's material, carries out keratocyte and cultivates.In this respect, Griffith in 1999 at United States Patent (USP) U.S6, disclosed " Artificial cornea " has 3 kinds of cells of cornea in 645,715, but is the immortalized cells through virus transfection, is not suitable for transplanting.It makes up cornea model is to use traditional keratocyte to mix with the collagen I gel and solidify the back generation, has shrinkage characteristics, the mechanical strength deficiency.Dimitriievich in 1999 at United States Patent (USP) U.S6; 471; Disclosed in 958 " Non-contracting tissue equivalent " also is that keratocyte mixes with collagen I type gel and solidifies the back and make up corneal stroma; Thereafter make up corneal epithelium and endodermis respectively, mix the shrinkage problems after solidifying though solved traditional keratocyte with collagen I type gel, transparency is not enough.
People seek the animal corneal of suitable implant into body in the animal of heterogenous allosome for many years; Trial is obtained animal corneal from animals such as cattle, pig, chicken, rabbit, fish, monkeys, handles to slough through a series of biochemistrys to cause that human immunity repels histiocyte and albumen makes it to fit and the human cornea transplanting.The subject matter of the method existence at present is that matter was opaque firmly, implants to produce immunological rejection, causes cornea tissue blood vessel hyperplasia and intraocular inflammation, causes the corneal transplantation failure after animal corneal passed through a series of biochemistrys processing.
Summary of the invention
To the shortcoming and deficiency of prior art, primary and foremost purpose of the present invention is to provide a kind of good biocompatibility, high, the good active regeneration artificial cornea graft that it is easy to make of transparency of matrix strength.
Another object of the present invention is to provide a kind of simple and be easy to the method for preparing of the above-mentioned corneal graft of suitability for industrialized production.
The object of the invention is realized through following technical scheme: a kind of active regeneration artificial cornea graft comprises the component of following weight portion:
Chitosan: 5～30 parts;
Collagen: 5～30 parts;
Chondroitin sulfate: 0.1～0.5 part;
Polyethylene Glycol: 5～10 parts;
Polyvinyl alcohol: 5～25 parts;
Tri-distilled water: 15～80 parts.
For making the present invention have more performance:
Also comprise keratocyte somatomedin, fibronectin and laminin in the component, their final mass concentration in raw mix is respectively: 10ng/ml～120ng/ml, 1ng/ml～100ng/ml and 1ng/ml～100ng/ml; Wherein the keratocyte somatomedin comprises in the middle of fibroblast growth factor (bFGF), transforming growth factor (TGF-β 1) and epidermal growth factor (EGF) or they two or more mixture.
In addition, in the said components: the molecular weight of Polyethylene Glycol preferred 1000～50000; The molecular weight of polyvinyl alcohol preferred 10000～100000.
The present invention provides a kind of method for preparing of active regeneration artificial cornea graft, comprises following concrete steps:
(1) take by weighing 5～30 parts of chitosans, 5～30 parts of collagen be dissolved in respectively pH 3 hydrochloric acid solution or acetum, 0.1～0.5 part of chondroitin sulfate is dissolved in tri-distilled water;
(2) sneak into 5～10 parts of Polyethylene Glycol again, 5～25 parts of polyvinyl alcohol and 15～80 parts of tri-distilled waters at room temperature stir 1～2hr, make the abundant mix homogeneously of each component;
(3) mixed liquor is injected mould, dry or place the freezer dryer lyophilizing;
(4) be soaked in water and clean obtain product;
Above umber is a weight portion.
Method step can be added keratocyte somatomedin, fibronectin and laminin in (2), and their mass concentration is respectively: 10ng/ml～120ng/ml, 1ng/ml～100ng/ml and 1ng/ml～100ng/ml.
Above-mentioned keratocyte somatomedin comprises fibroblast growth factor (bFGF), transforming growth factor (TGF-β
1) and epidermal growth factor (EGF) or they in the middle of two or more mixture; Preferred 10～the 100ng/ml of its consumption.
The molecular weight of Polyethylene Glycol is 1000～50000 in the above-mentioned steps (2), and the molecular weight of polyvinyl alcohol is 10000～100000;
The programmed cooling scope of freezing dry process is in the above-mentioned steps (3): 0～-120 degree, cooling process 0--10--20--30--40--50--60--70--80--90--100--110--120 degree.
The method for preparing of above-mentioned any active regeneration artificial cornea graft, the product in the step (4) can further be processed loose structure through laser boring.
The aperture of above-mentioned laser boring is 1～300 μ m, and pass is lines groove, circle, ellipse or fusiformis, and pitch-row is 100～1000 μ m.
Action principle of the present invention is:
In the component that corneal graft of the present invention comprised; Collagen is 75% the composition that accounts for people's cornea dry weight; It is the structural protein of people's keratocyte epimatrix; Have that good biocompatibility, biological degradability are good, no antigen, can promote advantages such as keratocyte growth, but collagen degradation in vivo speed is fast, mechanical performance is relatively poor, and its function as the keratocyte support is restricted.
Polyvinyl alcohol in the component (PVA) is a kind of water miscible polyhydroxylated polymer, has good water-solubility, nontoxic, the most important thing is that it has with the human body natural to organize close water content, elastic modelling quantity, low-friction coefficient and higher mechanical strength, abundant hole network structure, excellent biological compatibility; The character of PVA changes with relative molecular mass and the different of alcoholysis degree; Good film-forming property, intensity is high, appearance transparent; In body not by degraded by collagenase, the scalable degradation rate.Therefore, through changing its changes of properties of PVA relative molecular mass and alcoholysis degree regulation and control.Compound polyvinyl alcohol and collagen, both can improve the shortcoming of the bad mechanical property of collagen, can strengthen the cellular affinity of polyvinyl alcohol again.
Adhesion, the growing multiplication of the chondroitin sulfate corneal endotheliocyte in the component have good effect, and the scalable cell adhesion has the good cell affinity.It is to constitute outside the keratocyte and the main component of intercellular matrix; Have viscoelasticity and macromole hydrability; Can promote and the growth and breeding of pilot angle theca cell that RF corneal form and the keeping of normal physiological state that forms that combine with collagen has important function, through with the combining of corneal endothelium; Form the protecting film on cornea tissue surface, the damage that the radical pair keratocyte causes in the control volume.
In optimized technical scheme of the present invention, add 3 kinds of keratocyte somatomedin (fibroblast growth factor (bFGF), transforming growth factor (TGF-β simultaneously
1) with epidermal growth factor (EGF)), integrate cellulose fiber and be connected albumen and laminin.The cooperative effect between keratocyte somatomedin and the integration element and the adhesion and the growth and breeding of consumption corneal cell can be regulated and control; Promote between keratocyte normal fusion and with the combining of active regeneration artificial cornea graft, help the repair and reconstruction of impaired pathological changes cornea.
The three-dimensional porous stereochemical structure of active regeneration artificial cornea graft is the key factor that influences its performance.The present invention adopts lyophilization, Polyethylene Glycol and laser boring pore, makes up three-dimensional porous active regeneration artificial cornea graft.Polyethylene Glycol is a kind of nontoxic high molecular polymer, with water fabulous intersolubility is arranged, and its relative molecular mass can from several thousand to several ten thousand.Relative molecular mass size and the consumption of control PEG, the size of adjustable orifice, volume and density.Logical loose structure can be grown cell by rack surface in support, forming to internal migration.Control cryodesiccated temperature range, programmed cooling and each set of dispense ratio, size, volume and density that also can adjustment hole.In addition, adopt laser boring,, design different passes, aperture and pitch-row, can obtain more good three-dimensional porous stereochemical structure through control laser energy size.
The present invention compared with prior art has following beneficial effect:
(1) active regeneration artificial cornea graft that obtains of the present invention can solve that the conea equivalent seed cell is not suitable for transplanting, problems such as matrix strength deficiency and contraction, poor transparency, artificial cornea are leaked, front and back film formation.
(2) moisture content 60%～95% of the active regeneration artificial cornea graft of the present invention's acquisition, expansion rate 100%～500%, light transmittance 90%～100%, tensile strength (hygrometric state) 3～15Mpa, elongation at break 300%～500%, index of refraction 1.360～1.390.And the performance of existing same type of material is generally: material light transmittance 90%～96%, material refractive index 1.368～1.382 (eye cornea is 1.377); Mechanical strength and eye cornea are similar, wherein hygrometric state tensile strength 4Mpa～8.6MPa; Moisture content 55%～75%; Elongation at break 150%～300%;
(3) active regeneration artificial cornea that obtains of the present invention is suitable for keratocyte and grows in vivo and in vitro, but the pilot angle theca cell therein 3 D stereo adhere to growth; The speed of degraded and absorbed and the keratocyte speed of growth are complementary in vivo and in vitro;
(4) the present invention adopts lyophilization, Polyethylene Glycol and laser boring pore, makes up three-dimensional porous active regeneration artificial cornea graft.Use laser boring,, design different passes, aperture and pitch-row through control laser energy size; Control size, volume and density that cryodesiccated temperature range, programmed cooling and each set of dispense can adjustment holes than also; The relative molecular mass size and the consumption of control Polyethylene Glycol, the size of adjustable orifice, volume and density, logical loose structure in support, forming.
(5) prepare active regeneration artificial cornea graft according to bionic principle, its component proportioning adopts natural substrates collagen, chondroitin sulfate, chitosan to integrate keratocyte somatomedin (fibroblast growth factor (bFGF), transforming growth factor (TGF-β
1) with epidermal growth factor (EGF)), integrate cellulose fiber and be connected albumen and laminin.According to the adjustable component proportioning of different keratocytes; The cooperative effect between keratocyte somatomedin and the integration element and the adhesion and the growth and breeding of consumption corneal cell can be regulated and control; Promote between keratocyte normal fusion and with the combining of active regeneration artificial cornea graft, help the repair and reconstruction of impaired pathological changes cornea.
(6) active regeneration artificial cornea graft of the present invention can be processed mouldingly, can prepare the goods of different curvature and thickness, and forming method is easy to operate, and raw material is easy to get inexpensive, can carry out large-scale industrialized production.
The specific embodiment:
Embodiment 1
By weight or mass concentration, active regeneration artificial cornea graft comprises following component
Chitosan: 10 parts; Collagen: 10 parts;
Chondroitin sulfate: 0.3 part; Tri-distilled water: 67 parts;
Polyethylene Glycol (molecular weight 1000): 5 parts;
Polyvinyl alcohol (molecular weight 10000): 10 parts;
Keratocyte somatomedin: 10ng/ml;
Fibronectin: 60ng/ml;
Laminin: 25ng/ml;
Wherein the keratocyte somatomedin consists of fibroblast growth factor (bFGF) 10ng/ml.
Above-mentioned active regeneration artificial cornea graft is prepared by following steps: (1) takes by weighing chitosan, collagen and chondroitin sulfate according to the above ratio, with chitosan, collagen be dissolved in respectively pH 3 hydrochloric acid solution or acetum, chondroitin sulfate is dissolved in tri-distilled water; (2) mix above-mentioned each component, at room temperature stir 1～2hr, make the abundant mix homogeneously of each component; (3) mixed liquor being injected mould dries; (4) be soaked in water and obtain goods behind the cleaning, drying; (5) resulting product is used laser boring, aperture 150 μ m, pass lines groove, pitch-row 500 μ m.
The active regeneration artificial cornea graft that the present invention obtains: moisture content 60%, expansion rate 100%, light transmittance 90%, tensile strength (hygrometric state) 3Mpa, elongation at break 300%, index of refraction 1.360.
Embodiment 2
By weight or mass concentration, active regeneration artificial cornea graft comprises following component:
Chitosan: 20 parts; Collagen: 25 parts;
Chondroitin sulfate: 0.2 part; Tri-distilled water: 45 parts;
Polyethylene Glycol (molecular weight 4000): 5 parts;
Polyvinyl alcohol (molecular weight 40000): 5 parts;
Keratocyte somatomedin: 65ng/ml;
Fibronectin: 70ng/ml;
Laminin: 25ng/ml;
Wherein the keratocyte somatomedin consists of fibroblast growth factor (bFGF) 20ng/ml, epidermal growth factor (EGF) 30ng/ml, transforming growth factor (TGF-β
1) 15ng/ml.
Above-mentioned active regeneration artificial cornea graft is prepared by following steps: (1) takes by weighing chitosan, collagen and chondroitin sulfate according to the above ratio, with chitosan, collagen be dissolved in respectively pH 3 hydrochloric acid solution or acetum, chondroitin sulfate is dissolved in tri-distilled water; (2) mix above-mentioned each component, at room temperature stir 1～2hr, make the abundant mix homogeneously of each component; (3) mixed liquor being injected mould dries; (4) be soaked in water and obtain goods behind the cleaning, drying; (5) resulting product is used laser boring, aperture 300 μ m, and pass is circular, pitch-row 500 μ m.
The moisture content 95% of the active regeneration artificial cornea graft that the present invention obtains, expansion rate 500%, light transmittance 100%, tensile strength (hygrometric state) 15Mpa, elongation at break 500%, index of refraction 1.390.
Embodiment 3
By weight or mass concentration, active regeneration artificial cornea graft comprises following component:
Chitosan: 30 parts; Collagen: 15 parts;
Chondroitin sulfate: 0.4 part; Tri-distilled water: 37 parts;
Polyethylene Glycol (molecular weight 50000): 5 parts;
Polyvinyl alcohol (molecular weight 100000): 5 parts;
Keratocyte somatomedin: 75ng/ml;
Fibronectin: 40ng/ml;
Laminin: 55ng/ml;
Wherein the keratocyte somatomedin consists of fibroblast growth factor (bFGF) 30ng/ml, epidermal growth factor (EGF) 20ng/ml, transforming growth factor (TGF-β
1) 25ng/ml.
Above-mentioned active regeneration artificial cornea graft is prepared by following steps: (1) takes by weighing chitosan, collagen and chondroitin sulfate according to the above ratio, with chitosan be dissolved in pH 3 hydrochloric acid solution, collagen be dissolved in pH 3 acetum, chondroitin sulfate is dissolved in tri-distilled water; (2) mix above-mentioned each component, at room temperature stir 1～2hr, make the abundant mix homogeneously of each component; (3) mixed liquor being injected mould dries; (4) be soaked in water and obtain goods behind the cleaning, drying; (5) resulting product is used laser boring, aperture 100 μ m, and pass is oval, pitch-row 1000 μ m.
The moisture content 75% of the active regeneration artificial cornea graft that the present invention obtains, expansion rate 180%, light transmittance 98%, tensile strength (hygrometric state) 11Mpa, elongation at break 380%, index of refraction 1.380.
Embodiment 4
By weight or mass concentration, active regeneration artificial cornea graft comprises following component:
Chitosan: 15 parts; Collagen: 25 parts;
Chondroitin sulfate: 0.1 part; Tri-distilled water: 50 parts;
Polyethylene Glycol (molecular weight 20000): 5 parts;
Polyvinyl alcohol (molecular weight 20000): 5 parts;
Keratocyte somatomedin: 90ng/ml;
Dimension connects albumen: 50ng/ml;
Laminin: 35ng/ml;
Wherein the keratocyte somatomedin consists of fibroblast growth factor (bFGF) 30ng/ml, epidermal growth factor (EGF) 30ng/ml, transforming growth factor (TGF-β
1) 30ng/ml.
Above-mentioned active regeneration artificial cornea graft is prepared by following steps: (1) takes by weighing chitosan, collagen and chondroitin sulfate according to the above ratio, with chitosan, collagen be dissolved in respectively pH 3 hydrochloric acid solution, chondroitin sulfate is water-soluble; (2) mix above-mentioned each component, at room temperature stir 1～2hr, make the abundant mix homogeneously of each component; (3) mixed liquor is injected mould, place the freezer dryer lyophilizing, lyophilization and temperature programming temperature range :-100～-20 degree (4) are soaked in water and obtain goods behind the cleaning, drying;
The moisture content 70% of the active regeneration artificial cornea graft that the present invention obtains, expansion rate 150%, light transmittance 93%, tensile strength (hygrometric state) 13Mpa, elongation at break 350%, index of refraction 1.368.
Embodiment 5
By weight or mass concentration, active regeneration artificial cornea graft comprises following component:
Chitosan: 5 parts; Collagen: 30 parts;
Chondroitin sulfate: 0.5 part; Tri-distilled water: 80 parts;
Polyethylene Glycol (molecular weight 4000): 10 parts;
Polyvinyl alcohol (molecular weight 60000): 25 parts;
Keratocyte somatomedin: 120ng/ml;
Fibronectin: 100ng/ml;
Laminin: 120ng/ml;
Wherein the keratocyte somatomedin consists of fibroblast growth factor (bFGF) 50ng/ml, epidermal growth factor (EGF) 40ng/ml, transforming growth factor (TGF-β
1) 30ng/ml.
Above-mentioned active regeneration artificial cornea graft is prepared by following steps: (1) takes by weighing chitosan, collagen and chondroitin sulfate according to the above ratio, with chitosan be dissolved in pH 3 hydrochloric acid solution, collagen be dissolved in pH 3 acetum, chondroitin sulfate is water-soluble; (2) mix above-mentioned each component, at room temperature stir 1～2hr, make the abundant mix homogeneously of each component; (3) mixed liquor being injected mould dries; (4) be soaked in water and obtain goods behind the cleaning, drying; (5) resulting product is used laser boring, aperture 1 μ m, pass shuttle type, pitch-row 100 μ m.
The moisture content 76% of the active regeneration artificial cornea graft that the present invention obtains, expansion rate 330%, light transmittance 93%, tensile strength (hygrometric state) 8Mpa, elongation at break 380%, index of refraction 1.377.
Embodiment 6
By weight or mass concentration, active regeneration artificial cornea graft comprises following component:
Chitosan: 30 parts; Collagen: 5 parts;
Chondroitin sulfate: 0.1 part; Tri-distilled water: 15 parts;
Polyethylene Glycol (molecular weight 3000): 5 parts;
Polyvinyl alcohol (molecular weight 80000): 5 parts;
Keratocyte somatomedin: 50ng/ml;
Fibronectin: 1ng/ml;
Laminin: 1ng/ml;
Wherein the keratocyte somatomedin consists of epidermal growth factor (EGF) 50ng/ml.
Above-mentioned active regeneration artificial cornea graft is prepared by following steps: (1) takes by weighing chitosan, collagen and chondroitin sulfate according to the above ratio, with chitosan be dissolved in pH 3 hydrochloric acid solution, collagen be dissolved in pH 3 acetum, chondroitin sulfate is water-soluble; (2) mix above-mentioned each component, at room temperature stir 1～2hr, make the abundant mix homogeneously of each component; (3) mixed liquor being injected mould dries; (4) be soaked in water and obtain goods behind the cleaning, drying; (5) resulting product is used laser boring, aperture 300 μ m, and pass is oval, pitch-row 1000 μ m.
The moisture content 85% of the active regeneration artificial cornea graft that the present invention obtains, expansion rate 200%, light transmittance 98%, tensile strength (hygrometric state) 8.5Mpa, elongation at break 400%, index of refraction 1.373.
The foregoing description is a preferred implementation of the present invention; But embodiment of the present invention is not restricted to the described embodiments; Other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; All should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (8)
1. active regeneration artificial cornea graft is characterized in that comprising the component of following weight portion:
Chitosan: 5～30 parts
Collagen: 5～30 parts
Chondroitin sulfate: 0.1～0.5 part
Polyethylene Glycol: 5～10 parts
Polyvinyl alcohol: 5～25 parts
Tri-distilled water: 15～80 parts,
Also comprise keratocyte somatomedin, fibronectin and laminin, their mass concentration is respectively: 10ng/ml～120ng/ml, 1ng/ml～100ng/ml and 1ng/ml～100ng/ml;
Described active regeneration artificial cornea graft is prepared by following steps:
(1) take by weighing hydrochloric acid solution or the acetum that 5～30 parts of chitosans, 5～30 parts of collagens are dissolved in pH＜3 respectively, 0.1～0.5 part of chondroitin sulfate is dissolved in tri-distilled water;
(2) sneak into 5～10 parts of Polyethylene Glycol again, 5～25 parts of polyvinyl alcohol, and add tri-distilled water to 15～80 part, and at room temperature stir 1～2hr, make the abundant mix homogeneously of each component;
(3) mixed liquor is injected mould, dry or place the freezer dryer lyophilizing;
(4) be soaked in water and clean obtain product;
Above umber is a weight portion;
Add keratocyte somatomedin, fibronectin and laminin in the said step (2), their mass concentration is respectively: 10ng/ml～120ng/ml, 1ng/ml～100ng/ml and 1ng/ml～100ng/ml.
2. according to the said active regeneration artificial cornea graft of claim 1, it is characterized in that: said keratocyte somatomedin is fibroblast growth factor bFGF, epidermal growth factor TGF-β 1, transforming growth factor EGF or two or more mixture in the middle of them.
3. according to the said active regeneration artificial cornea graft of claim 1, it is characterized in that: the molecular weight of said Polyethylene Glycol is 1000～50000; The molecular weight of said polyvinyl alcohol is 10000～100000.
4. the method for preparing of the said active regeneration artificial cornea graft of claim 1 is characterized in that comprising following concrete steps:
(1) take by weighing hydrochloric acid solution or the acetum that 5～30 parts of chitosans, 5～30 parts of collagens are dissolved in pH＜3 respectively, 0.1～0.5 part of chondroitin sulfate is dissolved in tri-distilled water;
(2) sneak into 5～10 parts of Polyethylene Glycol again, 5～25 parts of polyvinyl alcohol, and add tri-distilled water to 15～80 part, and at room temperature stir 1～2hr, make the abundant mix homogeneously of each component;
(3) mixed liquor is injected mould, dry or place the freezer dryer lyophilizing;
(4) be soaked in water and clean obtain product;
Above umber is a weight portion;
Add keratocyte somatomedin, fibronectin and laminin in the said step (2), their mass concentration is respectively: 10ng/ml～120ng/ml, 1ng/ml～100ng/ml and 1ng/ml～100ng/ml.
5. according to the method for preparing of the said active regeneration artificial cornea graft of claim 4, it is characterized in that: said keratocyte somatomedin comprises fibroblast growth factor bFGF, epidermal growth factor TGF-β 1, transforming growth factor EGF or two or more mixture in the middle of them.
6. according to the method for preparing of the said active regeneration artificial cornea graft of claim 4, it is characterized in that: the molecular weight of Polyethylene Glycol is 1000～50000 in the said step (2), and the molecular weight of polyvinyl alcohol is 10000～100000; The programmed cooling scope of freezing dry process is in the said step (3): 0～-120 degree, cooling process 0--10--20--30--40--50--60--70--80--90--100--110--120 degree.
7. according to the method for preparing of each said active regeneration artificial cornea graft of claim 4～6, it is characterized in that: the product in the said step (4) is processed loose structure through laser boring.
8. according to the method for preparing of the said active regeneration artificial cornea graft of claim 7, it is characterized in that: the aperture of said laser boring is 1～300 μ m, and pass is lines groove, circle, ellipse or fusiformis, and pitch-row is 100～1000 μ m.
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| CN101632841B (en) * | 2009-08-21 | 2012-10-03 | 暨南大学 | Tissue engineering scaffold containing alginate, glycosaminoglycan and collagen and preparation method thereof |
| CN101947144B (en) * | 2010-09-29 | 2012-07-04 | 厦门大学 | Ply tissue engineering corneal frame and manufacturing method and application thereof |
| CN105013010B (en) * | 2015-07-07 | 2018-01-12 | 中山大学中山眼科中心 | A kind of laminin film of auxiliary iPS RPE transplanting |
| 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 |
| CN106798949B (en) * | 2017-02-22 | 2019-09-20 | 福建吉特瑞生物科技有限公司 | A kind of porous hydroxyapatite bone renovating material and preparation method thereof |
| CN107412861B (en) * | 2017-02-24 | 2021-08-06 | 武汉采思生物科技有限公司 | Bone repair gel of recombinant collagen compounded with chondroitin sulfate and polyethylene glycol |
| CN112370571A (en) * | 2020-11-26 | 2021-02-19 | 同济大学 | Bone tissue engineering scaffold material for treating jaw and face bone defect and synthesis method thereof |
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