CN1256990C - Preparation method of plant origion alcohrl soluble protein three dimentional support - Google Patents
Preparation method of plant origion alcohrl soluble protein three dimentional support Download PDFInfo
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- CN1256990C CN1256990C CN 200410015674 CN200410015674A CN1256990C CN 1256990 C CN1256990 C CN 1256990C CN 200410015674 CN200410015674 CN 200410015674 CN 200410015674 A CN200410015674 A CN 200410015674A CN 1256990 C CN1256990 C CN 1256990C
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Abstract
The present invention relates to a preparation method of a three-dimensional support made from plant-derived alcohol soluble protein, which belongs to the technical field of biology. The preparation method comprises: using the plant-derived alcohol soluble protein as a main component and mixing with a pore-forming agent according to a mass ratio of 1: 0 to 1: 10, or independently using the protein and naturally forming at 10 to 95 DEG C in a high moisture environment, or utilizing a mould to extrude a prearranged shape; choosing common salt, ammonium bicarbonate, ammonium acetate, ammonia chloride and benzamide as the pore-forming agent; mixing with a synthetic polymer of polylactic acid and polyglycollic acid, or hybridizing with known biologic medical materials comprising collagen, chitosan and hydroxyapatite. Vitro cell culture experiments of the present invention all show that the protein has favorable cell compatibility and material characteristics required by tissue engineering, and thus, a novel biologic medical material with wide sources, degradability, low immunogenicity, microorganism resistance and proper mechanical strength is found for the tissue engineering.
Description
Technical field
The present invention relates to a kind of preparation method of plant source protein matter, particularly a kind of preparation method of Phytogenous alcohol-soluble protein three-dimensional rack.Belong to biological technical field.
Background technology
The reparation of the development of modern medicine, particularly tissue and organ or the development of implantation technique and popularize become a big market to the demand of bio-medical material.To the requirement of bio-medical material, except that having excellent biological compatibility, nontoxic, degradable in vivo, also to have certain mechanical strength.Be the microcellular structure body especially for what the material of tissue engineering bracket also required certain intensity, cell can be sticked on surface and inside; Help the growth and the breeding of cell; Help the exchange of cell desired nutritional material and the eliminating of metabolic waste.The organic substance that is used for bio-medical material at present can roughly be divided into synthetic high polymer such as polylactic acid etc. and natural polymer.There has been the natural polymer of report that protein is arranged, polysaccharide etc., collagen for example, chitosan.And proteinic kind is very limited.Synthetic high polymer has the good mechanical processing characteristics, but its biocompatibility is not ideal enough; The advantage of natural polymer is a degradable, good biocompatibility, but mechanical strength is not enough, thereby their application is restricted.
Zein was identified out in 1897, was the main protein in the corn, accounted for the 45%-50% of its total protein, can be processed into resin, had toughness, hydrophobicity, degradability, antibiotic property etc.At present, zein has obtained commercial applications.In medical treatment, the adhesive, the binder that have Showa etc. to produce, the medicament carrier microspheres of design such as Mathiowitz, the odor mask of the oral drugs of Cuca invention; The cosmetics aspect, the face powder that has Avalle to formulate; The packaging material for food of developing as OSAK and Wasa etc. on the food, the food antioxidant of invention such as king.In addition, the application of zein also relates to all many-sides such as chewing gum, paint, pad-ink, film, degradability plastics, hair dye, optical fiber and textile.But do not see the relevant report that is applied to bio-medical material.
We have at first imagined the potential using value of this protein as bio-medical material, to its biocompatibility, inside and outside degradability, mechanics processing characteristics, three-dimensional rack process technology (Wang Jinye etc., application number 02138761.3), two-dimensional film technology of preparing CN 1401659A such as () Wang Jinye, microsphere preparation technology application numbers 03129639.4 such as () Wang Jinye that contains bioactive molecule and disinfecting technical etc. have carried out system, deep research, all obtain satisfactory result.Prove that it is the new type natural bio-medical material that a kind of utmost point has development potentiality really.Particularly can obtain the support of suitable mechanical strength, be better than any natural macromolecular material of having reported, and can satisfy the high requirement of bone tissue engineer mechanical property by suitable method.Though said method can access the timbering material with higher compression intensity of tens of MPa, porosity is lower, and manufacturing cycle is longer.These problems to be solved by this invention just.
Summary of the invention
The objective of the invention is to overcome deficiency of the prior art, a kind of preparation method of Phytogenous alcohol-soluble protein three-dimensional rack is provided.Making it is that the bio-medical material of main component preparation can reach more than 70% with the porosity of three-dimensional porous rack with the Phytogenous alcohol-soluble protein, and method of the present invention can be met the three-dimensional porous rack of organizational project needs in short process time.Be a kind of degradable, have higher mechanical strength, the biocompatibility excellent material.
The present invention is achieved by the following technical solutions, and the present invention is to be main component with the Phytogenous alcohol-soluble protein, presses the different quality ratio with the pore former of variable grain size, protein: pore former=1: 0-1: 10, mix, perhaps use protein separately, utilize mould to be pressed into predetermined shape.
Use 60-100 ℃ of (recommended temperature is 85-95C) decocting in water 10 minutes-8 hours (the recommendation time is 3-5 hour) then, make crosslinked tightr between the protein.Sample for adding pore former will change water 3-5 time, to remove pore former, vacuum lyophilization then fully in the decocting in water process.
Except that Phytogenous alcohol-soluble protein is main component, also contain variable grain size and ratio as known pore formers such as Sal, Ammonium bicarbonate food grade, ammonium acetate, ammonia chloride, benzoic acid amine.
Except that Phytogenous alcohol-soluble protein is main component, also mix synthetic high polymers such as polylactic acid, polyglycolic acid, perhaps hybridize with collagen protein, chitosan, the known bio-medical material of hydroxyapatite.
Described plant source protein is a kind of deriving from as corn, Semen Tritici aestivi, Fructus Hordei Vulgaris, naked barley, avenaceous protein,alcohol-soluble.
The present invention has substantial characteristics and obvious improvement, after process the inventive method is carried out forming processes, can obtain having higher mechanical strength, and aperture and porosity are adjustable, degradable three-dimensional porous rack.In cell culture experiments in vitro, mensuration by morphological observation, cell viability, adhesive force proves that all this albumen has the good cell compatibility, possessed the desired material characteristics of organizational project, thereby for organizational project has found a kind of wide material sources, degradable, reduced immunogenicity, antibiotic property, new bio medical material with suitable mechanical strength.Phytogenous alcohol-soluble protein stroma not only help cell adherent, stretch and growth, and can be processed into three-dimensional porous rack, and under parody pendular ring border, can degrade, what is more important can satisfy organizational project, even the desired mechanical strength of Os Elephatis tissue engineering material.Its raw material is easy to get, and preparation method is easy, is a kind of coming new bio medical material.
Description of drawings
The growth rate (mtt assay) of Fig. 1 .NIH3T3 on different materials.
Fig. 2. the degradation curve (n=4) of zein support in the citricacid-Na2HPO4 of pH 2.2 buffer.
Fig. 3. the sem photograph of the support that the present invention is prepared (a is the enlarged drawing of b, and the particulate diameter of Sal is the 76-150 micron).
Specific embodiments
Phytogenous alcohol-soluble protein support of the present invention, help cell adherent, stretch and growth, as being seeded in three-dimensional rack after last 2 hour with the NIH3T3 mouse fibroblast cell, almost all be attached on the support, cell viability (90%) is compared with matched group (75%) greatly to be increased, in the proliferate afterwards, the one-tenth fiber characteristics of cell is (seeing accompanying drawing 1) especially obviously.This support can be degraded under parody pendular ring border, and degradation speed can be regulated (seeing accompanying drawing 2) by the pore-forming condition.Can obtain the three-dimensional rack of mechanics modulus of compressibility with the method for the invention in hundreds of KPa-40Mpa scopes; The aperture is-700 microns of tens of microns; Porosity is at 50%-98%, and can regulate.
Embodiment one
With zein with after pore former mixes as stated above, pack in the cylindrical die according to the proteic amount of each sample 1 gram, take out after the pressure compression forming with about 800N, each sample is placed on separately in the small beaker that fills the 60C deionized water, to prevent sample adhesion mutually in water, direct decocting in water 1 hour in thermostat water bath changes pre-heated deionized water 2-3 time, to guarantee that Sal is dissolved out fully in the decocting in water process then.Lyophilisation then, measuring the modulus of compressibility modulus is 10MPa.The scanning electron microscope Ultrastructural observation shows that the aperture is distributed as (Fig. 3) about 300 microns.
Claims (3)
1, a kind of preparation method of Phytogenous alcohol-soluble protein three-dimensional rack, it is characterized in that, with the Phytogenous alcohol-soluble protein is main component, press mass ratio with pore former, protein: pore former=1: 0-1: 10, mix, perhaps use protein separately, utilize mould to be pressed into predetermined shape, reuse 60-100 ℃ decocting in water 10 minutes-8 hours makes crosslinked tightr between the protein, changes pre-heated deionized water in the decocting in water process 2-3 time, to guarantee to remove fully pore former, vacuum lyophilization then.
2, the preparation method of Phytogenous alcohol-soluble protein three-dimensional rack according to claim 1, described plant source protein are a kind of corn, Semen Tritici aestivi, Fructus Hordei Vulgaris, naked barley or avenaceous protein,alcohol-solubles of deriving from.
3, the preparation method of Phytogenous alcohol-soluble protein three-dimensional rack according to claim 1, described pore former comprises: Sal, Ammonium bicarbonate food grade, ammonium acetate, ammonia chloride, benzoic acid amine.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200410015674 CN1256990C (en) | 2004-01-08 | 2004-01-08 | Preparation method of plant origion alcohrl soluble protein three dimentional support |
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| CN 200410015674 CN1256990C (en) | 2004-01-08 | 2004-01-08 | Preparation method of plant origion alcohrl soluble protein three dimentional support |
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| CN1555892A CN1555892A (en) | 2004-12-22 |
| CN1256990C true CN1256990C (en) | 2006-05-24 |
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100336565C (en) * | 2005-12-01 | 2007-09-12 | 上海交通大学 | Plant source protein three-dimensional stent material |
| CN100349623C (en) * | 2006-04-27 | 2007-11-21 | 上海交通大学 | Active gliadin porous stent preparation method |
| CN103948961B (en) * | 2014-04-28 | 2016-06-08 | 上海交通大学 | Corn protein materials, Preparation method and use |
| CN105664247A (en) * | 2014-11-19 | 2016-06-15 | 中国科学院上海应用物理研究所 | Nanometer calcium silicate fiber/corn protein composite material as well as preparation method and applications of nanometer calcium silicate fiber/corn protein composite material |
| CN104721885A (en) * | 2015-03-06 | 2015-06-24 | 青岛大学附属医院 | Mesoporous calcium magnesium silicate/wheat protein composite material as well as preparation method and application of composite material |
| US20220154018A1 (en) * | 2019-01-08 | 2022-05-19 | Shanghai Jiao Tong University | A bioink for 3d printing, the preparation method and usage |
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