CN102908666A - Collagen scaffold material for cornea - Google Patents
Collagen scaffold material for cornea Download PDFInfo
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- CN102908666A CN102908666A CN2012103824998A CN201210382499A CN102908666A CN 102908666 A CN102908666 A CN 102908666A CN 2012103824998 A CN2012103824998 A CN 2012103824998A CN 201210382499 A CN201210382499 A CN 201210382499A CN 102908666 A CN102908666 A CN 102908666A
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Abstract
The invention discloses a scaffold material for clinical cornea reconstruction and a preparation method of the material, belongs to the field of medical biomaterials, and mainly aims at solving the present problems that the clinical supply of cornea donors is difficult and the success rate of operations is low. The cornea is reconstructed by providing a keratocyte growth matrix for a cornea receptor. The material prepared by the method is prepared by adjusting pH values of polyose and a collagen solution to be within a certain range at a certain environmental temperature and then conducting crosslinking membrane forming. The material is stable, uniform, transparent, water-proof and pressure-resistant, and is good in biocompatibility. The cornea reconstruction of the corneal transplantation receptor is facilitated.
Description
Technical field
The present invention relates to composite of a kind of polysaccharide and collagen and preparation method thereof.This material has certain light transmission, and outward appearance becomes film like.The keratocyte that is used for before the ophthalmology cornea is transplanted is cultivated support.Belong to the biomaterial for medical purpose field.
Background technology
10 million people's blind is to be caused by keratopathy in the world at present, for causing the second largest reason of blind, is only second to cataract.Wherein 80% can cast off illiteracy by corneal graft.Yet traditional corneal graft has its two large drawback, the one, and the source difficulty of cornea donor, the 2nd, success rate of operation is low.
In view of this, develop particularly important that the front timbering material of breeding for the peculiar cornea cells survival of patient of a kind of keratoplasty becomes.Again because the structure mesostroma layer 90% of natural ocular cornea is collagen, therefore our company has adopted the method for utilizing polysaccharide and collagen to carry out structural remodeling to prepare this type of compound support frame material.The material that the method makes, evenly, printing opacity, certain crushing resistance is arranged, waterproof, the chief component composition has good biocompatibility near natural cornea tissue.Can cultivate for the keratocyte of corneal transplantation receptor good support is provided.
Summary of the invention
This material selection polysaccharide and the collagen method of carrying out structural remodeling make.Can satisfy keratocyte and cultivate the requirement that reaches cornea reconstruction.Main operational steps is as follows:
1, the preparation of collagen solution: one or more of commercially available type i collagen, II Collagen Type VI, III Collagen Type VI, be dissolved in the hydrochloric acid of 0.01mol/L～0.5mol/L, stir, make the solution that concentration is 0.1%～15.0% (w/w).
2, compounding ingredients preparation: one or both polysaccharide in commercially available chitosan hydrochlorate, using carboxyl chitosan, glycosaminoglycans, Dan Baiduotang proteoglycan PG, chitosan lactate, chitosan quaternary amine, chitosan sulfate fat, hyaluronic acid-like chitosan, sodium alginate, hyaluronic acid, sodium carboxymethyl cellulose, hydroxy ethyl fiber sodium or the HPMC and the collagen in the collagen solution are (1～10) in mass ratio: the ratio of (10～1) mixes.
3, aqueous slkali: commercially available sodium hydroxide, solution preparation, making its concentration is 0.01mol/L～5.0mol/L.
4, the preparation of collagen-based cornea timbering material: compounding ingredients is mix and blend under the environment of (1 ℃～40 ℃) in temperature, and behind 0.5h～24h, the aqueous slkali adjust pH makes final pH value be (6～8).At last gained solution is carried out the crosslinked film forming of bed board.
The specific embodiment
Embodiment 1,
Get commercially available chitosan hydrochlorate 1.0g, slowly pour 100ml concentration into and be in 5.0% the collagen solution, pour into while stirring.Wherein collagen solution is that the mixing collagen of type i collagen and III Collagen Type VI is dissolved in 0.2mol/L hydrochloric acid.After stirring, stirred 23 ℃ of ambient temperatures 1 hour.With 1.0mol/L sodium hydroxide solution adjust pH to 7.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 1: 5;
Embodiment 2,
Get commercially available sodium alginate 1.3g, slowly pour 100ml concentration into and be in 13.0% the collagen solution, pour into while stirring.Wherein collagen solution is that the mixing collagen of type i collagen and II Collagen Type VI is dissolved in 3.0mol/L hydrochloric acid.After stirring, stirred 23 ℃ of ambient temperatures 7 hours.With 5.0mol/L sodium hydroxide solution adjust pH to 6.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 1: 10.
Embodiment 3,
Get commercially available using carboxyl chitosan 1.5g, slowly pour 100ml concentration into and be in 1.0% the collagen solution, pour into while stirring.Wherein collagen solution is that the mixing collagen of type i collagen and III Collagen Type VI is dissolved in 0.2mol/L hydrochloric acid.Stir and stirred 38 ℃ of ambient temperatures 12 hours.With 1.0mol/L sodium hydroxide solution adjust pH to 7.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 3: 2.
Embodiment 4,
Get commercially available chitosan lactate 5.0g, slowly pour 100ml concentration into and be in 5.0% the collagen solution, pour into while stirring.Wherein collagen solution is that the mixing collagen of type i collagen and II Collagen Type VI and III Collagen Type VI is dissolved in 0.2mol/L hydrochloric acid.After stirring, stirred 2 ℃ of ambient temperatures 8 hours.With 3.0mol/L sodium hydroxide solution adjust pH to 6.8.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 1: 1.
Embodiment 5,
Get commercially available chitosan quaternary amine 10.0g, slowly pour 100ml concentration into and be in 10.0% the collagen solution, pour into while stirring.Wherein collagen solution is that the mixing collagen of II Collagen Type VI and III Collagen Type VI is dissolved in 0.2mol/L hydrochloric acid.After stirring, stirred 10 ℃ of ambient temperatures 2 hours.With 0.5mol/L sodium hydroxide solution adjust pH to 7.3.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 1: 1.
Embodiment 6,
Get commercially available chitosan sulfate fat 2.0g, slowly pour 100ml concentration into and be in 0.5% the collagen solution, pour into while stirring.Collagen solution is that type i collagen is dissolved in 0.2mol/L hydrochloric acid, after stirring, stirs 15 ℃ of ambient temperatures 0.5 hour.With 2.5mol/L sodium hydroxide solution adjust pH to 7.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 4: 1.
Embodiment 7,
Get commercially available hyaluronic acid-like chitosan 3.0g, slowly pour 100ml concentration into and be in 2.0% the collagen solution, pour into while stirring.Wherein collagen solution is that the mixing collagen of type i collagen and III Collagen Type VI is dissolved in 0.2mol/L hydrochloric acid, after stirring, stirs 18 ℃ of ambient temperatures 5 hours.With 2.8mol/L sodium hydroxide solution adjust pH to 6.5.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 3: 2.
Embodiment 8,
Get commercially available sodium carboxymethyl cellulose 8.0g, slowly pour 100ml concentration into and be in 2.0% the collagen solution, pour into while stirring.Wherein collagen solution is that the mixing collagen of II Collagen Type VI and III Collagen Type VI is dissolved in 0.2mol/L hydrochloric acid, after stirring, stirs 1 ℃ of ambient temperature 10 hours.With 4.0mol/L sodium hydroxide solution adjust pH to 8.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 4: 1.
Embodiment 9,
Get commercially available chitosan hydrochlorate 1.5g and sodium carboxymethyl cellulose 1.5g, slowly pour respectively 100ml concentration into and be in 1.5% the collagen solution, pour into while stirring.Wherein collagen solution is that type i collagen is dissolved in 1.0mol/L hydrochloric acid, after stirring, stirs 25 ℃ of ambient temperatures 18 hours.With 2.0mol/L sodium hydroxide solution adjust pH to 8.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 2: 1.
Embodiment 10,
Get commercially available hyaluronic acid 3.0g, slowly pour 100ml concentration into and be in 2.0% the collagen solution, pour into while stirring.Wherein collagen solution is that II Collagen Type VI and III Collagen Type VI mixing collagen are dissolved in 0.2mol/L hydrochloric acid, after stirring, stirs 25 ℃ of ambient temperatures 16 hours.Make final pH value to 7.8 with 2.0mol/L sodium hydroxide solution adjust pH.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 3: 2.
Embodiment 11,
Get commercially available hydroxy ethyl fiber sodium 4.0g, slowly pour 100ml concentration into and be in 2.0% the collagen solution, pour into while stirring.Wherein collagen solution is that the II Collagen Type VI is dissolved in 0.2mol/L hydrochloric acid, after stirring, stirs 15 ℃ of ambient temperatures 4 hours.With 2.0mol/L sodium hydroxide solution adjust pH to 6.3.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 2: 1.
Embodiment 12,
Get commercially available hydroxy propyl cellulose sodium 4.2g, slowly pour 100ml concentration into and be in 1.8% the collagen solution, pour into while stirring.Wherein collagen solution is that the III Collagen Type VI is dissolved in 0.2mol/L hydrochloric acid, after stirring, stirs 8 ℃ of ambient temperatures 4 hours.0.8mol/L sodium hydroxide solution adjust pH to 8.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 7: 3.
Embodiment 13,
Get commercially available chitosan hydrochlorate 2.5g and hydroxy propyl cellulose sodium 2.0g, slowly pour respectively 100ml concentration into and be in 1.5% the collagen solution, pour into while stirring.Wherein collagen solution is that type i collagen is dissolved in 1.0mol/L hydrochloric acid.After stirring, stirred 35 ℃ of ambient temperatures 8 hours.With 3.8mol/L sodium hydroxide solution adjust pH to 6.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 3: 1.
Embodiment 14,
Get commercially available chitosan polysaccharide 10.5g, slowly pour 100ml concentration into and be in 1.5% the collagen solution, pour into while stirring.Wherein collagen solution is that type i collagen is dissolved in 1.0mol/L hydrochloric acid.Stir, stir 30 ℃ of 12 hours ambient temperatures.Transfer pH value to 6 with the 1.8mol/L sodium hydroxide solution.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 7: 1.
Embodiment 15,
Get commercially available glutelin polysaccharide 3.0g, slowly pour 100ml concentration into and be in 4.5% the collagen solution, pour into while stirring.Collagen solution is that type i collagen is dissolved in 1.0mol/L hydrochloric acid.After stirring, stirred 23 ℃ of ambient temperatures 24 hours.With 0.8mol/L sodium hydroxide solution adjust pH to 7.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 2: 3.
Embodiment 16,
Get commercially available chitosan hydrochlorate 1.2g and sodium alginate 1.5g, slowly pour respectively 100ml concentration into and be in 1.5% the collagen solution, pour into while stirring.Wherein collagen solution is that type i collagen is dissolved in 1.0mol/L hydrochloric acid.The rear stirring 20 hours that stirs, 23 ℃ of ambient temperatures.With 2.0mol/L sodium hydroxide solution adjust pH to 7.5.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 9: 5.
Embodiment 17,
Get commercially available sodium alginate 0.5g and hydroxy propyl cellulose sodium 2.0g, slowly pour respectively 100ml concentration into and be in 1.5% the collagen solution, pour into while stirring.Wherein collagen solution is that type i collagen is dissolved in 1.0mol/L hydrochloric acid.After stirring, stirred 5 ℃ of ambient temperatures 8 hours.With 2.8mol/L sodium hydroxide solution adjust pH to 6.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 5: 3.
Embodiment 18,
Get commercially available chitosan hydrochlorate 1.5g and sodium carboxymethyl cellulose 1.5g, slowly pour respectively 100ml concentration into and be in 1.5% the collagen solution, pour into while stirring.Wherein collagen solution is that type i collagen is dissolved in 0.5mol/L hydrochloric acid, after stirring, stirs 25 ℃ of ambient temperatures 18 hours.With 2.0mol/L sodium hydroxide solution adjust pH to 8.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 2: 1.
Embodiment 19,
Get commercially available chitosan hydrochlorate 8.0g, slowly pour 100ml concentration into and be in 10.0% the collagen solution, pour into while stirring.Wherein collagen solution is that type i collagen is dissolved in 0.01mol/L hydrochloric acid.After stirring, stirred 23 ℃ of ambient temperatures 20 hours.With 2.0mol/L sodium hydroxide solution adjust pH to 7.5.Gained composite bed board is cross-linked into film.The mass ratio of polysaccharide and collagen is 4: 5.
Claims (1)
1. the compound support frame material by polysaccharide and collagen is that polysaccharide and collagen solution mix the composite that carries out structural remodeling and form, and is cross-linked into film before packing.Its feature comprises the following steps:
(1), commercially available type i collagen, II Collagen Type VI, III Collagen Type VI one or more, be dissolved in the hydrochloric acid of 0.01mol/L～0.5mol/L, stir, make the solution that concentration is 0.1%～15.0% (w/w).
(2), commercially available chitosan hydrochlorate, using carboxyl chitosan, glycosaminoglycans, Dan Baiduotang proteoglycan PG, chitosan lactate, chitosan quaternary amine, chitosan sulfate fat, hyaluronic acid-like chitosan, sodium alginate, hyaluronic acid, sodium carboxymethyl cellulose, hydroxy ethyl fiber sodium or HPMC one or both polysaccharide and the collagen in the collagen solution wherein is (1～10) in mass ratio: the ratio of (10～1) mixes, and forms compounding ingredients.
(3), compounding ingredients is mix and blend under the environment of (1 ℃～40 ℃) in temperature, behind 0.5h～24h, the aqueous slkali adjust pH makes final pH value be (6～8).At last gained solution is carried out the crosslinked film forming of bed board.
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Cited By (6)
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| CN104383599A (en) * | 2014-11-05 | 2015-03-04 | 天津市赛瑞生物技术有限公司 | Collagen-based cornea scaffold and preparation method thereof |
| CN105833344A (en) * | 2016-04-26 | 2016-08-10 | 青岛慧生惠众生物科技有限公司 | Application of injectable hydrogel in preparing intraocular filling materials |
| 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 |
| CN111187443A (en) * | 2020-03-17 | 2020-05-22 | 陕西科技大学 | Collagen-sodium humate compound suitable for hemostatic material and preparation method thereof |
| CN112899215A (en) * | 2017-11-30 | 2021-06-04 | 山西加乐医疗科技有限责任公司 | Cell scaffold and preparation method thereof |
| CN113133535A (en) * | 2021-04-28 | 2021-07-20 | 华南理工大学 | Soluble non-denatured type II collagen-polysaccharide complex with digestion resistance and preparation method thereof |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104383599A (en) * | 2014-11-05 | 2015-03-04 | 天津市赛瑞生物技术有限公司 | Collagen-based cornea scaffold and preparation method thereof |
| CN105833344A (en) * | 2016-04-26 | 2016-08-10 | 青岛慧生惠众生物科技有限公司 | Application of injectable hydrogel in preparing intraocular filling materials |
| CN105833344B (en) * | 2016-04-26 | 2019-04-26 | 青岛慧生惠众生物科技有限公司 | A kind of injection aquagel is preparing the application in Ocular tamponades |
| 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 |
| CN112899215A (en) * | 2017-11-30 | 2021-06-04 | 山西加乐医疗科技有限责任公司 | Cell scaffold and preparation method thereof |
| CN111187443A (en) * | 2020-03-17 | 2020-05-22 | 陕西科技大学 | Collagen-sodium humate compound suitable for hemostatic material and preparation method thereof |
| CN113133535A (en) * | 2021-04-28 | 2021-07-20 | 华南理工大学 | Soluble non-denatured type II collagen-polysaccharide complex with digestion resistance and preparation method thereof |
| CN113133535B (en) * | 2021-04-28 | 2022-10-18 | 华南理工大学 | Soluble non-denatured type II collagen-polysaccharide complex with digestion resistance and preparation method thereof |
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Application publication date: 20130206 |