CN1382425A - Biologically induced artificial active cornea and its preparing process - Google Patents
Biologically induced artificial active cornea and its preparing process Download PDFInfo
- Publication number
- CN1382425A CN1382425A CN 02115215 CN02115215A CN1382425A CN 1382425 A CN1382425 A CN 1382425A CN 02115215 CN02115215 CN 02115215 CN 02115215 A CN02115215 A CN 02115215A CN 1382425 A CN1382425 A CN 1382425A
- Authority
- CN
- China
- Prior art keywords
- cornea
- induction type
- type active
- artificial cornea
- biological induction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Abstract
A biological induced artificial active cornea is prepared from chitosan (0.5-3 wt%), collagen (0.5-3 wt.%), hyaluronic acid (0.1-0.5 wt.%) and water (93.5-96 wt%), through dissolving chitosan in acid solution, dissolving collagen and hyaluronic acid in water, proportional mixing, moulding while baking, demoulding, and immersing and washing in water. Its advantages are high compatibility to eye tissue, and high performance similar to that of human cornea.
Description
(1) technical field
The present invention relates to artificial cornea's technology, particularly a kind of biological induction type active artificial cornea and preparation method thereof.
(2) background technology
Keratopathy is the present principal element of eye blinding, and global patient has 1,000 ten thousand people approximately, China 3,000,000 corneal blindness patients that also have an appointment.But because the scarcity of donor source, the allogeneic keratoplasty that China can carry out every year only is about 1000 examples, and a large amount of patients can not get curing because of the shortage of human cornea, and this is a global long-term unsolved problem.Because the no blood vessel characteristic of eye immune privilege phenomenon and cornea, keratoplasty is the highest operation of success rate in body tissue's organ transplantation, and therefore, artificial cornea and transplanting thereof are better than the transplanting of other organizational project organs and tissue.Present existing artificial cornea has two kinds of nonactive artificial cornea and the epigamic tissue engineering artificial corneals of inanimate object, the artificial cornea that nonactive artificial cornea utilizes synthetic material such as methyl methacrylate (MMA) etc. to make, when carrying out the human implantation, though can make people's vision recovers to some extent, but field range is minimum, synthetic material is difficult to merge with enclosing near the eyes to organize mutually, foreign body sensation is arranged after the implantation, corneal solution, leakage of aqueous humor appear easily, the artificial cornea deviates from and complication such as endophthalmitis, brings very big misery to the patient.And common notional tissue engineering artificial corneal is under the condition of external reinforcement, carries out keratocyte and cultivates.The cell of In vitro culture just makes it biologically active after growth on the timbering material; Though the artificial cornea that this method is produced may make moderate progress aspect the histocompatibility with enclosing near the eyes, but because it needs the multiple keratocyte of In vitro culture, and the approaches and methods of acquisition of keratocyte at present and cultivation is relatively more difficult and immature, the preservation after keratocyte is cultivated and the also existing problems of breeding that go down to posterity, whole process is complicated, expense is high, and the immunologic rejection effect of allosome keratocyte in migration process also is a great problem that faces.
(3) summary of the invention
The objective of the invention is to overcome the shortcoming of prior art, have excellent biological compatibility, easy to use, functional and long preservation and long-distance transporting but provide a kind of with people ocular tissue, the biological induction type active artificial cornea of being convenient to carry out large-scale production and realizing industrialization.
Another object of the present invention is to provide a kind of method for preparing above-mentioned biological induction type active artificial cornea.
Purpose of the present invention is achieved through the following technical solutions: this biological induction type active artificial cornea comprises following component---
Chitosan 0.5%～3%
Collagen 0.5%～3%
Hyaluronic acid 0.1%～0.5%
Water 93.5%～96.0%
Above ratio is weight percentage.
Except that above-mentioned component, for helping stimulate speed from the enrichment of the bulk-growth factor, this biological induction type active artificial cornea also can be integrated with the keratocyte somatomedin as becoming very thin somatomedin (FGF), epithelical cell growth factor (EGF) and cell transformation somatomedin (TGF-β
1), consumption is 3～100ng/ml, the carbodiimide with 0.01%～0.2% (EDC) cross-linking agent is crosslinked, forms the biological induction type active artificial cornea of band somatomedin.
The method for preparing above-mentioned biological induction type active artificial cornea comprises the steps: that (1) is dissolved in acid solution with chitosan, and collagen and hyaluronic acid are water-soluble; (2) mix each component; (3) mixed liquor is injected mould and oven dry; (4) be soaked in water behind cleaning, drying resulting product and make it reach the suction balance.
Behind each component mix homogeneously of step 2, can add the keratocyte somatomedin.
Described acid solution is hydrochloric acid solution or acetum.
Described mixed process makes the abundant mix homogeneously of each component at room temperature stirring 1～2hr.
Action principle of the present invention is: people's cornea tissue mainly is made of epithelial cell, fibroblast, endotheliocyte and collagen fiber and mucopolysaccharide, collagen protein accounts for 75% of cornea dry weight, can from animal, extract in a large number, the source is abundant and the charge is small, therefore adopts the host material of collagen as this artificial CF.
The collagen that from animal, extracts, though similar to human collagen on forming, variation has taken place in its overall structure in leaching process, the collagen after the purification has lost intensity, as the backing substrate material, is subjected to the effect of collagenase in vivo, very easily degraded; In order to remedy these weakness of collagen, strengthen intensity and its degradation rate in vivo of control of collagen, take to add the method for chitosan; Chitosan is a kind of natural biological aminopolysaccharide, to the mucopolysaccharide in the matter between keratocyte similar structure is arranged, good film-forming property, intensity height, appearance transparent, in body not by degraded by collagenase, can be degraded by lysozyme lentamente, thereby can regulate the degradation rate of backing substrate material, it is crosslinked also can to use cross-linking agent that material is carried out simultaneously, the control degradation rate.
Hyaluronic acid belongs to mucopolysaccharide; it is to constitute outside the keratocyte and the main component of intercellular matrix; have viscoelasticity and macromole hydrability; can promote growth and breeding with the pilot angle theca cell; network structure corneal form and the keeping of normal physiological state that forms that combine with collagen has important function; by 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; the hydroxyl that contains in the mucopolysaccharide composition on a large amount of electronegative carboxyls and eye surface in the hyaluronan molecule is easy to and the surperficial affinity of eye by hydrogen bonded.
The cultivation of somatomedin corneal cell, growth and breeding reach the great influence that is combined with extracellular and intercellular matrix, the cooperative effect between the different keratocyte somatomedin and the growth and breeding of consumption corneal cell can be regulated and control, wherein fibroblast growth factor (FGF), transforming growth factor (TGF-β
1), and the growth and breeding of epidermal growth factor (EGF) corneal cell, intercellular normal fusion and facilitation is all arranged with combining of substrate.
The present invention has following advantage with respect to prior art: (1) biological induction type active artificial cornea of the present invention and ocular tissue have the good compatibility, merge mutually with the cornea cell, induce the enrichment of multiple keratocyte somatomedin, stimulate keratocyte growth and breeding thereon; Before corneal transplantation and repairing, need not carry out the cultivation of external keratocyte, thereby exempted the complex process of In vitro culture keratocyte and transplanted the immunological rejection cause by the allosome keratocyte, and the various complication that produce therefrom, so use not only simple and convenient, and as safe as a house reliable, action effect is better; (2) this biological induction type active artificial cornea has optics similar to eye cornea and mechanical characteristic, can carry out various machine-shapings according to relevant requirements; The optics mechanical index of the biological induction type active artificial cornea of having made is similar to people's cornea, as follows: 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: 300%～350%; (3) this biological induction type active artificial cornea relatively is fit to processing and forming, and manufacturing process is simple, handling ease is convenient, so can prepare in large quantity and use; (4) this biological induction type active artificial cornea is easy to long preservation and long-distance transporting, and is convenient to carry out large-scale production and realizes industrialization; (5) invention of this biological induction type active artificial cornea, not only important facilitation is arranged for people ' s health, can produce important society and economy benefit, but also open up road for the research of other biological induction type active artificial organ, thereby have important scientific meaning.
(4) specific embodiment
Below in conjunction with embodiment the present invention is done further concrete description, but embodiments of the present invention are not limited thereto.
Embodiment 1
The ratio of each component of this biological induction type active artificial cornea is as follows:
Chitosan 2%
Collagen 2.5%
Hyaluronic acid 0.3%
Water 95.1%
Above ratio is weight percentage.
The process for preparing this biological induction type active artificial cornea with above-mentioned each component comprises the steps: that (1) takes by weighing each component by above-mentioned consumption, and chitosan is dissolved in hydrochloric acid solution, and collagen and hyaluronic acid is water-soluble; (2) chitosan salt acid solution and collagen and hyaluronic aqueous solution were at room temperature stirred 1.5 hours, make the abundant mix homogeneously of each component; (3) the carbodiimide cross-linking agent of adding 0.1% in the mixed liquor of mix homogeneously, 90ng/ml becomes very thin somatomedin, and 35ng/ml epithelical cell growth factor and 60ng/ml cell transformation somatomedin fully mix half an hour again; (4) mixed liquor is injected ready-formed cornea mould, fill with and take out after putting it into drying in oven behind the cornea mould; (5) with the cornea goods of clear water soaking and washing oven dry back gained removing residual impurity, and make it in water, place 24～48 hours to reach the suction balance, take out then and promptly make this biological induction type active artificial cornea.
Embodiment 2
The ratio of each component of this biological induction type active artificial cornea is as follows:
Chitosan 1%
Collagen 3%
Hyaluronic acid 0.2%
Water 95.65%
Above ratio is weight percentage.
The process for preparing this biological induction type active artificial cornea with above-mentioned each component comprises the steps: that (1) takes by weighing each component by above-mentioned consumption, and chitosan is dissolved in acetum, and collagen and hyaluronic acid is water-soluble; (2) chitosan-acetic acid solution and collagen and hyaluronic aqueous solution were at room temperature stirred 2 hours, make the abundant mix homogeneously of each component; (3) the carbodiimide cross-linking agent of adding 0.15% in the mixed liquor of mix homogeneously, 5ng/ml becomes very thin somatomedin, and 20ng/ml epithelical cell growth factor and 10ng/ml cell transformation somatomedin fully mix half an hour again; (4) mixed liquor is injected ready-formed cornea mould, fill with and take out after putting it into drying in oven behind the cornea mould; (5) with the cornea goods of clear water soaking and washing oven dry back gained removing residual impurity, and make it in water, place 24～48 hours to reach the suction balance, take out then and promptly make this biological induction type active artificial cornea.
Embodiment 3
The ratio of each component of this biological induction type active artificial cornea is as follows:
Chitosan 0.5%
Collagen 1.5%
Hyaluronic acid 0.3%
Water 97.7%
Above ratio is weight percentage.
The process for preparing this biological induction type active artificial cornea with above-mentioned each component comprises the steps: that (1) takes by weighing each component by above-mentioned consumption, and chitosan is dissolved in acetum, and collagen and hyaluronic acid is water-soluble; (2) chitosan-acetic acid solution and collagen and hyaluronic aqueous solution were at room temperature stirred 2 hours, make the abundant mix homogeneously of each component; (3) mixed liquor is injected ready-formed cornea mould, fill with and take out after putting it into drying in oven behind the cornea mould; (4) with the cornea goods of clear water soaking and washing oven dry back gained removing residual impurity, and make it in water, place 24～48 hours to reach the suction balance, take out then and promptly make this biological induction type active artificial cornea.
Embodiment 4
The ratio of each component of this biological induction type active artificial cornea is as follows:
Chitosan 1.5%
Collagen 2%
Hyaluronic acid 0.1%
Water 96.4%
Above ratio is weight percentage.
The process for preparing this biological induction type active artificial cornea with above-mentioned each component comprises the steps: that (1) takes by weighing each component by above-mentioned consumption, and chitosan is dissolved in acetum, and collagen and hyaluronic acid is water-soluble; (2) chitosan-acetic acid solution and collagen and hyaluronic aqueous solution were at room temperature stirred 2 hours, make the abundant mix homogeneously of each component; (3) mixed liquor is injected ready-formed cornea mould, fill with and take out after putting it into drying in oven behind the cornea mould; (4) with the cornea goods of clear water soaking and washing oven dry back gained removing residual impurity, and make it in water, place 24～48 hours to reach the suction balance, take out then and promptly make this biological induction type active artificial cornea.
The parameter of measuring the optics mechanical index of the biological induction type active artificial cornea that the various embodiments described above make and eye cornea is more as shown in Table 1:
Table one
| The optical force mathematic(al) parameter | Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 | The eye cornea value |
| Material light transmittance (%) | ????93％ | ????96％ | ????95％ | ????92％ | ????95-98％ |
| The material refractive index | ????1.370 | ????1.376 | ????1.380 | ????1.373 | ????1.377 |
| Hygrometric state tensile strength MPa | ????3.7 | ????4.0 | ????4.2 | ????4.1 | ????3.8 |
| Moisture content (%) | ????76％ | ????68％ | ????65％ | ????73％ | ????70％ |
| Elongation at break (%) | ???300％ | ????330％ | ????350％ | ????320％ |
By last table comparative result as can be known, this biological induction type active artificial cornea is very approaching in performance parameter and eye cornea value, this shows, this biological induction type active artificial cornea is comparatively similar to eye cornea.
The above-mentioned biological induction type active artificial cornea that makes is carried out animal heeling-in experiment, artificial cornea's flaggy transplantation experiments respectively.Be can be observed by experimental result: implanting this artificial cornea of back can keep transparent always, does the pathology section after 60 days after the taking-up dyeing and observes under optical microscope, and visible cornea structure is normal, the NIP cell, epithelium does not have and thickens and attenuation, and essential layer does not have edema, and interior skin structure is normal; The biological induction type artificial cornea material of implanting can be compatible with cornea, and is bonding between cornea and the material, with cornea mutually adhesion the visible keratocyte growth in material place and have trend to the material internal growth; Two months degradation rates in vivo of material are about 30%.
Claims (6)
1, a kind of biological induction type active artificial cornea comprises following component:
Chitosan 0.5%～3%
Collagen 0.5%～3%
Hyaluronic acid 0.1%～0.5%
Water 93.5%～96.0%
Above ratio is weight percentage.
2, according to the described biological induction type active artificial cornea of claim 1, it is characterized in that: be integrated with the keratocyte somatomedin.
3, according to the described biological induction type active artificial cornea of claim 2, it is characterized in that: described keratocyte somatomedin is for becoming very thin somatomedin, epithelical cell growth factor and cell transformation somatomedin, consumption is respectively 3～100ng/ml, and is crosslinked with 0.01%～0.2% carbodiimide cross-linking agent.
4, a kind of preparation method of biological induction type active artificial cornea is characterized in that comprising the steps: that (1) is dissolved in acid solution with chitosan, and collagen and hyaluronic acid are water-soluble; (2) mix each component; (3) mixed liquor is injected mould and oven dry; (4) be soaked in water behind cleaning, drying resulting product and make it reach the suction balance.
5, according to the preparation method of the described biological induction type active artificial cornea of claim 4, it is characterized in that: behind each component mix homogeneously, add the keratocyte somatomedin.
6, according to the preparation method of the described biological induction type active artificial cornea of claim 4, it is characterized in that: described acid solution is hydrochloric acid solution or acetum.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02115215 CN1228096C (en) | 2002-05-14 | 2002-05-14 | Biologically induced artificial active cornea and its preparing process |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02115215 CN1228096C (en) | 2002-05-14 | 2002-05-14 | Biologically induced artificial active cornea and its preparing process |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1382425A true CN1382425A (en) | 2002-12-04 |
| CN1228096C CN1228096C (en) | 2005-11-23 |
Family
ID=4743517
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 02115215 Expired - Lifetime CN1228096C (en) | 2002-05-14 | 2002-05-14 | Biologically induced artificial active cornea and its preparing process |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1228096C (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1321704C (en) * | 2004-03-24 | 2007-06-20 | 华东理工大学 | Method for fabricating activated artificial skin tissue in bilayer by using bioreactor |
| CN100512889C (en) * | 2006-12-08 | 2009-07-15 | 华南理工大学 | Process of making cornea histoengineering support in bionic structure |
| CN101112625B (en) * | 2006-07-24 | 2010-09-01 | 陶鹭 | Chitose shield for treating ceratonosus and method of manufacturing the same |
| CN102188746A (en) * | 2010-03-11 | 2011-09-21 | 北京益而康生物工程开发中心 | Artificial extracellular matrix and preparation method thereof |
| CN103483625A (en) * | 2013-09-09 | 2014-01-01 | 戴建英 | Absorbable and degradable multipurpose biocompatible material |
| CN109907861A (en) * | 2019-04-22 | 2019-06-21 | 清华大学深圳研究生院 | A kind of multifunctionality corneal graft |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107185038A (en) * | 2017-05-19 | 2017-09-22 | 广州市朴道联信生物科技有限公司 | A kind of surface immobilized modified cornea repair material and preparation method thereof |
-
2002
- 2002-05-14 CN CN 02115215 patent/CN1228096C/en not_active Expired - Lifetime
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1321704C (en) * | 2004-03-24 | 2007-06-20 | 华东理工大学 | Method for fabricating activated artificial skin tissue in bilayer by using bioreactor |
| CN101112625B (en) * | 2006-07-24 | 2010-09-01 | 陶鹭 | Chitose shield for treating ceratonosus and method of manufacturing the same |
| CN100512889C (en) * | 2006-12-08 | 2009-07-15 | 华南理工大学 | Process of making cornea histoengineering support in bionic structure |
| CN102188746A (en) * | 2010-03-11 | 2011-09-21 | 北京益而康生物工程开发中心 | Artificial extracellular matrix and preparation method thereof |
| CN103483625A (en) * | 2013-09-09 | 2014-01-01 | 戴建英 | Absorbable and degradable multipurpose biocompatible material |
| CN103483625B (en) * | 2013-09-09 | 2015-09-23 | 戴建英 | The multi-usage biocompatible materials of absorbable and degradable |
| CN109907861A (en) * | 2019-04-22 | 2019-06-21 | 清华大学深圳研究生院 | A kind of multifunctionality corneal graft |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1228096C (en) | 2005-11-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101433478B (en) | Whole layer biological cornea as well as construction method and use thereof | |
| CN104436285B (en) | A kind of regenerated silk fibroin gel mould and preparation method thereof | |
| EP2755598B1 (en) | Fabrication of gelatin hydrogel sheet for the transplantation of corneal endothelium | |
| CN101302486B (en) | Acetobacter xylinum and method for preparing nano-cellulose skin tissue repair material by using the same | |
| CN110772669A (en) | Biological ink for 3D printing of artificial skin | |
| CN101380486B (en) | Active regeneration artificial cornea graft and preparation method thereof | |
| CN102886063A (en) | Preparation and application of cellulose nanocrystals (CNCs)-reinforced collagen compound substrate | |
| CN102807965B (en) | Method for preparing tissue engineered cornea and device of method | |
| CN1228096C (en) | Biologically induced artificial active cornea and its preparing process | |
| CN100512889C (en) | Process of making cornea histoengineering support in bionic structure | |
| US20180216148A1 (en) | Composite cellulose hydrogels and methods of making and use thereof | |
| CN111671973A (en) | Preparation method of polypyrrole/silk fibroin composite conductive tissue engineering scaffold | |
| CN1218755C (en) | Stem cell regenerating surface cornea and its application in corneal transplantation | |
| CN101543642B (en) | Collagen-based interpenetrating polymer network tissue engineering cornea substitute and preparation method thereof | |
| Mahdavi et al. | Development and in vitro evaluation of photocurable GelMA/PEGDA hybrid hydrogel for corneal stromal cells delivery | |
| CN101480505B (en) | Method for preparing surface-functionalized chitosan cornea repair material | |
| CN100340309C (en) | Biological induction type active artificial cornea, and application method therefor | |
| EP4011411A1 (en) | Preparation method for silk fibroin nerve graft fused with nt3 | |
| CN102166374B (en) | Method for preparing amniotic compound corneal limbus stem cell membrane | |
| CN113637067A (en) | Recombinant human collagen and artificial cornea thereof | |
| CN105148325A (en) | New cornea tissue repair material and preparing method thereof | |
| CN103239762B (en) | Chitin membrane and application of chitin membrane in ophthalmologic treatment | |
| CN113354837B (en) | Patterned inverse opal collagen hydrogel for in-vitro tissue scaffold construction and preparation method thereof | |
| CN104941001A (en) | Biological film corneal patch and preparation method and application thereof | |
| CN105963791B (en) | A kind of high bioactivity chitin timbering material and its preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CX01 | Expiry of patent term |
Granted publication date: 20051123 |
|
| CX01 | Expiry of patent term |