CN104001213A - Porous support for cartilage tissue engineering and preparation method thereof - Google Patents
Porous support for cartilage tissue engineering and preparation method thereof Download PDFInfo
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- CN104001213A CN104001213A CN201410190481.7A CN201410190481A CN104001213A CN 104001213 A CN104001213 A CN 104001213A CN 201410190481 A CN201410190481 A CN 201410190481A CN 104001213 A CN104001213 A CN 104001213A
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Abstract
The invention relates to a porous support for cartilage tissue engineering and a preparation method thereof, and belongs to the technical field of biological medical materials. The porous support is a three-dimensional porous support comprising carragheenan and chitosan and has porosity of 80-96% and aperture sizes of 50-500 microns. The preparation method comprises the following steps of preparing a carragheenan aqueous solution from distilled water, preparing a chitosan solution from a dilute acid solution, respectively beating the solutions, then blending the solutions, carrying out stirring, adding sodium carbonate crystal powder into the mixture, carrying out beating again, carrying out water-bath treatment, pouring the treated mixture into a stainless steel mold, carrying out freeze drying, carrying out immersion in a dilute acid solution, carrying out immersion in distilled water and carrying out vacuum drying to obtain the porous support for cartilage tissue engineering. The preparation method utilizes the raw material having wide sources and safety. The porous support has a certain function, high hardness, high porosity and no residue and is suitable for tissue cell culture or cartilage regeneration. The preparation method has simple and easy processes and a wide industrial prospect.
Description
Technical field
The present invention relates to bio-medical material technical field, specifically a kind of used in tissue engineering porous support and preparation method thereof.
Background technology
Carrageenan (carrageenan), has another name called carrageenin, is the natural anion polysaccharide plant colloid extracting from red algae.It can be divided into κ type carrageenan, ι type carrageenan and λ type carrageenan etc. according to the difference of structure type, best with the coagulability of κ type carrageenan.Carrageenan is that white is to khaki, surface glossy, translucent plates or Powdered thing, odorless, tasteless, mouthfeel stick-slip expands in cold water, dissolve in and form the transparent or slight milky easy fluent solution of viscosity after 60 ℃ of above hot water, be insoluble to organic solvent.The easy gel of carrageenan, has thermal reversibility, after heating for dissolving, can form semi-solid transparent gel while letting cool.Because the half ester formula sulfate group on its residue is with negative charge, makes it under the cationic inductions such as potassium, ammonium, calcium, sodium, to form double-stranded supermolecule bundle, thereby improve widely its gel strength.Under micro-potassium chloride (0.1%) exists, can improve the gel strength of 3 times, but too high gel strength can cause gel excessively crisp.0.5% sodium carbonate exists can improve the gel strength of carrageenan 70%, and can reduce carrageenan transparency, enhancing hardness.Carrageenan stability is strong, heat and also can not be hydrolyzed, but in acid solution (especially pH≤4.0), acid hydrolysis easily occurs carrageenan, gel strength and viscosity degradation in neutral and alkaline solution.This peculiar property of carrageenan makes it as thickening agent, gellant, suspending agent, emulsifying agent and stabilizing agent etc., be widely used in the fields such as food industry, chemical industry and biochemistry, medical research.In addition, carrageenan and derivant thereof are also proved has multiple biological activity, as antitumor, anticoagulation, enhancing immunity and antiviral etc.The application prospect of carrageenan is very considerable.
Chitosan (chitosan) is Chitosan, and another name chitosan, is the natural active matter being made by de-acetyl chitin, is extensively present in the animal bodies such as shrimp and crab shells, insecticide.Chitosan is white or faint yellow powder, stable in properties, and water insoluble and aqueous slkali, dissolves in diluted acid.Under sour environment, its side chain is amino can be positively charged in conjunction with proton.Chitosan has good film property, biocompatibility and biological activity, nontoxic, harmless, without immunizing antigen, and human body is had reinforced immunological, anti-bacteria, the recovery from illness of promotion disease, regulates the functions such as physiological function.Chitosan can be made into film and is applied to the aspects such as fruit freshness preserving film; Can also and be applied to medical aspect as flocculating agent, fiber, liquid crystal, catalyst, adsorbent; In food industry, can be used as antibacterial, edible film, food additive, sewage-treating agent, medicinal slow release agent, enzyme immobilization carrier etc.Its application prospect is tempting, and current study hotspot is the aspects such as medicament slow release that it has.
Organizational project (tissue engineering) is the philosophy and technique of application life sciences and engineering, in the mammiferous organizational structure and emic basis normally and under 2 kinds of states of pathology of correct understanding, research and development are for repairing, safeguard, promote a new branch of science of the biosubstitute of the function and morphology after the various tissues of human body or organ injury.Organizational project has tempting application prospect on clinical medicine.The core of Tissue Engineering Study is to set up the three-dimensional complex consisting of cell, biomaterial scaffolds and somatomedin now.Wherein, the cytoskeletal effect consisting of biomaterial is to provide space for cell proliferation, make cell under the effect of somatomedin, break up, breed according to the configuration of biomaterial scaffolds, finally become desired tissue or organ, reach the object of repairing wound and Reconstruction of The Function.Therefore, the preparation of three-dimensional porous rack is the precondition of engineered human organ.Current, the common used material of three-dimensional rack comprise natural macromolecular material as collagen, gelatin, cellulose, chitin, chitosan, alginate, glycosaminoglycan, agar, liposome etc. with inorganic material as light base apatite, calcium phosphate etc.In addition, also have some synthetic macromolecular materials, as collagen and polylactic acid, collagen and light base apatite, chitosan and to gather light base sour, collagen and chitosan and other polymer are such as gathering the compound etc. of phosphazo, polyester urethane.Different materials all has its different nature and extent, but about histiocyte cultivate and cartilage tissue regeneration class much all use gelatin or collagen as support.As everyone knows, current gelatin is faced with huge under-supply present situation, there are a lot of security incidents that flow into food and medicine market about gelatin inferior, therefore, be necessary to explore the three-dimensional porous rack material that more can be used for histiocyte cultivation and cartilage tissue regeneration class.About the technology of preparing of three-dimensional porous rack material, current have microsphere sintering process, solution-cast/particle leaching method, be separated/freeze-drying of thermic, method of electrostatic spinning, a supercritical fluid preparation method etc.More traditional microsphere sintering process pore rate is low; Solution-cast/particle leaching method easily causes dissolvent residual, and the solvent of volatilization is environmental protection not also; Be separated/freeze-drying of thermic is not accurate enough on control porosity and aperture; Method of electrostatic spinning, the contour technical method of supercritical fluid preparation method will have larger prospect really in future, but its current cost is too high, and extensive use is still unrealistic.
Crosslinked compound studies confirm that by multidigit scholar of carrageenan-chitosan, some has also prepared carrageenan-chitosan micropill, but as three-dimensional porous rack, still be there are to many problems in the cross-linked composite of carrageenan-chitosan, as carrageenan hardness is strong, gel porosity is not high, aperture is difficult to control etc.
Summary of the invention
In order to solve above prior art problem,
The object of the present invention is to provide a kind of cartilage tissue engineered porous support and preparation method thereof of using, this porous support hardness is higher, porosity is higher, and without hazard residue, safety, have necessarily functionally, can be used in that histiocyte is cultivated or cartilage tissue regeneration, preparation method is simply effective.
The technical solution adopted in the present invention is:
A kind of cartilage tissue engineered with porous support, described cartilage tissue engineered with porous support by carrageenan and chitosan by (8～4): the three-dimensional porous rack that 1 mass ratio forms, porosity is 80～96%, aperture is 50～500 μ m, modulus of compressibility 0.7～1.2MPa, stretch modulus 1.0～2.2MPa.
Preferably, described carrageenan is the mixture of κ type carrageenan κ type or κ type carrageenan, ι type carrageenan, λ type carrageenan, and mixing quality ratio is κ type carrageenan: ι type carrageenan: λ type carrageenan=(5～9): (1～5): the epoxy glue of (1～5).
Preferably, described chitosan is the chitosan that molecular weight is 100,000～500,000, deacetylation is greater than 80%.
Another object of the present invention is to provide a kind of described cartilage tissue engineered method with porous support of preparing, and said method comprising the steps of:
(1) by the carrageenan solutions of 60 ℃～70 ℃ distilled water preparations 3%～15%, with the chitosan solution of dilute acid soln preparation 0.5%～1.2%, the two is pulled an oar respectively;
(2) by above-mentioned carrageenan solutions and chitosan solution blend, stir, obtain carrageenan-chitosan complexes;
(3) by the making beating of carrageenan-chitosan complexes, add crystals of sodium carbonate powder, making beating again, obtains carrageenan-chitosan-carbon acid sodium complex;
(4) by carrageenan-chitosan-carbon acid sodium complex water-bath 20～50min at 60 ℃～70 ℃, be cast in mould, be put in freezing 3～6h at-40 ℃～-20 ℃, lyophilization, obtains carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket;
(5) carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket is immersed in dilute acid soln, soak 3～5h, then use distilled water immersion 8～10h, 2～4h changes water once, and vacuum drying, obtains cartilage tissue engineered with carrageenan-chitosan composite porous support afterwards.
Preferably, the described carrageenan solutions of step (2) and chitosan solution are according to carrageenan and chitosan solute mass ratio (8～4): 1 ratio blend.
Preferably, in step (3), crystals of sodium carbonate powder and carrageenan-chitosan complexes solute mass ratio are (6～15): 1.
Preferably, the size of the crystals of sodium carbonate powder described in step (3) is 100～200 μ m.
Preferably, hydrochloric acid or acetic acid that the described dilute acid soln of step (1) is PH3～4, carrageenan and chitosan are cross-linked compound in left and right, PH3～4, and Material cladding is more even; Hydrochloric acid or acetic acid that the described diluted acid of step (5) is pH1.5～2.5.
Preferably, the described pulping process of step (1) adopts beater, and beating rate is 8000～10000r/min, and beating time is 20～30s; Magnetic stirring apparatus is used in the described stirring of step (2), and mixing speed is 1000～2000r/min.
Preferably, the described lyophilization of step (4) is vacuum 10～20Pa, 0 ℃ of lower pumping 5～10h; The described vacuum drying of step (5) is vacuum 10～20Pa 5～10h that bleeds.
The present invention with respect to the beneficial effect of prior art is:
1) the present invention selects with the carrageenan solutions of anion and with cationic chitosan solution, is cross-linked compoundly, generates carrageenan-chitosan complexes and prepares three-dimensional rack.Do not use gelatin, natural material wide material sources used, and there is certain biological activity, can there be good biocompatibility, degradability.
2) the present invention selects carrageenan and chitosan as the raw material of porous support, wide material sources, and safety, has necessarily functional;
3) after material dissolution of the present invention, process respectively through making beating, can make crosslinked compound more fully, the product that gel structure stability is timbering material than available technology adopting gelatin or collagen improves 40%;
4) the present invention is cross-linked compound by carrageenan and chitosan under low sour acid condition, because carrageenan does not have gelation under acidity, so Material cladding is even, the material that is support than existing gelatin or collagen-chitin, the uniformity improves 30%;
5) the present invention adds a large amount of crystals of sodium carbonate powder to carry out pore, the in the situation that of neutralization part acid ion, produces gas, increase hole, and low acidity of the present invention can guarantee that sodium carbonate can be by too much decomposition; After partial sodium carbonate is decomposed, PH rises, and some sodium carbonate are dissolved in hole, have increased gel strength and the hardness of carrageenan gel, also have some still with crystal state, to be present in the hole of carrageenan-chitosan complexes, the porosity of maintenance complex can be lower than 80%; When lyophilization, the sodium carbonate of dissolving has part and separates out increase porosity; The aperture of preparation method medium-height trestle of the present invention is easily controlled;
6) the present invention can effectively prepare the three-dimensional porous rack that hardness is higher, modulus of compressibility reaches 0.7～1.2Mpa, stretch modulus reaches 1.0～2.2MPa and porosity is high, and do not have hazard residue, carrageenan-chitosan complexes can be used in histiocyte and cultivates or cartilage tissue regeneration, novel unique, have a extensive future.
Accompanying drawing explanation
Fig. 1 is the cartilage tissue engineered SEM photo with porous support in the embodiment of the present invention;
Hematoxylin-Yihong colored graph of rabbit joint implant frame cambium after 12 weeks in Fig. 2 embodiment 3;
The sarranine dyeing mucopolysaccharide figure of rabbit joint implant frame cambium after 12 weeks in Fig. 3 embodiment 3.
The specific embodiment
Below by embodiment, the present invention is described in further details, these embodiment are only used for illustrating the present invention, do not limit the scope of the invention.
Embodiment 1
A kind of cartilage tissue engineered with porous support, the three-dimensional porous rack being formed by the mass ratio of 6:1 by the chitosan of κ type carrageenan and molecular weight 100,000, deacetylation 90%, porosity 89%, aperture 100～150 μ m, modulus of compressibility 0.7MPa, stretch modulus 1.0MPa.
A cartilage tissue engineered preparation method with porous support, comprises the following steps:
(1) by the carrageenan solutions of 60 ℃ of distilled water preparations 3%, the chitosan solution with PH3 dilute hydrochloric acid solution preparation 0.6%, is used respectively beater 8000r/min by it, making beating 30s;
(2) by above-mentioned carrageenan and chitosan solution according to carrageenan and chitosan solute mass ratio 6:1 blend, use magnetic stirring apparatus 1000r/min to stir, obtain carrageenan-chitosan complexes;
(3) by the making beating of carrageenan-chitosan complexes, add 120 μ m crystals of sodium carbonate powder (with solute mass ratio in carrageenan-chitosan complexes be 10:1), making beating again, obtains carrageenan-chitosan-carbon acid sodium complex;
(4) by 70 ℃ of water-bath 30min of carrageenan-chitosan-carbon acid sodium complex, in casting stainless steel mould, be put in freezing 4h at-30 ℃, vacuum 10Pa, 0 ℃ of lower pumping 6h lyophilization, obtain carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket;
(5) carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket is immersed in the hydrochloric acid solution of pH1.5, soak 3h, then use distilled water immersion 8h, 2h changes water once, the 10Pa 7h vacuum drying of bleeding, obtains cartilage tissue engineered with carrageenan-chitosan composite porous support afterwards.
Embodiment 2
A kind of cartilage tissue engineered with porous support, the three-dimensional porous rack being formed by the mass ratio of 4:1 by the chitosan of κ type carrageenan and molecular weight 200,000, deacetylation 85%, porosity 92%, aperture 50～80 μ m, modulus of compressibility 1.2MPa, stretch modulus 2.2MPa.
A cartilage tissue engineered preparation method with porous support, comprises the following steps:
(1) by the carrageenan solutions of 65 ℃ of distilled water preparations 5%, the chitosan solution with PH3.6 acetic acid,diluted solution preparation 1%, is used respectively beater 8500r/min by it, making beating 25s;
(2) by above-mentioned carrageenan and chitosan solution according to carrageenan and chitosan solute mass ratio 5:1 blend, use magnetic stirring apparatus 1500r/min to stir, obtain carrageenan-chitosan complexes;
(3) by the making beating of carrageenan-chitosan complexes, add 100 μ m crystals of sodium carbonate powder (with solute mass ratio in carrageenan-chitosan complexes be 6:1), making beating again, obtains carrageenan-chitosan-carbon acid sodium complex;
(4) by 60 ℃ of water-bath 50min of carrageenan-chitosan-carbon acid sodium complex, in casting stainless steel mould, be put in freezing 6h at-20 ℃, vacuum 15Pa, 0 ℃ of lower pumping 7h lyophilization, obtain carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket;
(5) carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket is immersed in the acetic acid solution of pH2, soak 5h, then use distilled water immersion 9h, 3h changes water once, the 15Pa 6h vacuum drying of bleeding, obtains cartilage tissue engineered with carrageenan-chitosan composite porous support afterwards.
Embodiment 3
A kind of cartilage tissue engineered with porous support, by the epoxy glue of κ type carrageenan, ι type carrageenan and λ type carrageenan, (mixing quality ratio is κ type carrageenan: ι type carrageenan: three-dimensional porous rack λ type carrageenan=8:1:1) forming by the mass ratio of 8:1 with the chitosan of molecular weight 330,000, deacetylation 92%, porosity 90%, aperture 200～260 μ m, modulus of compressibility 1.0MPa, stretch modulus 2.0MPa.
A cartilage tissue engineered preparation method with porous support, comprises the following steps:
(1) by the carrageenan solutions of 67 ℃ of distilled water preparations 10%, the chitosan solution with PH3.3 dilute hydrochloric acid solution preparation 0.5%, is used respectively beater 10000r/min by it, making beating 25s;
(2) by above-mentioned carrageenan and chitosan solution according to carrageenan and chitosan solute mass ratio 8:1 blend, use magnetic stirring apparatus 2000r/min to stir, obtain carrageenan-chitosan complexes;
(3) by the making beating of carrageenan-chitosan complexes, add 170 μ m crystals of sodium carbonate powder (with solute mass ratio in carrageenan-chitosan complexes be 9:1), making beating again, obtains carrageenan-chitosan-carbon acid sodium complex;
(4) by 67 ℃ of water-bath 40min of carrageenan-chitosan-carbon acid sodium complex, in casting stainless steel mould, be put in freezing 5h at-33 ℃, vacuum 20Pa, 0 ℃ of lower pumping 9h lyophilization, obtain carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket;
(5) carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket is immersed in the acetic acid solution of pH1.5, soak 5h, then use distilled water immersion 9h, 2h changes water once, the 20Pa 9h vacuum drying of bleeding, obtains cartilage tissue engineered with carrageenan-chitosan composite porous support afterwards.
(6) compound rest step (5) being obtained is planted new zealand white rabbit knee joint osteochondral defect place, and sampling gross examination of skeletal muscle is as figure after 12 weeks, and visible damaged place is smooth smooth, and cambium fills up whole damaged and be combined with host tissue well; By 12 weeks samples in body, carry out respectively hematoxylin-Yihong dyeing (Fig. 2), mucopolysaccharide dyeing (Fig. 3), can see that neocartilage is rich in mucopolysaccharide, thickness and autologous cartilage thickness are basically identical, and are combined well with newborn subchondral bone.
Embodiment 4
A kind of used in tissue engineering carrageenan-chitosan composite porous support, by the epoxy glue of κ type carrageenan, ι type carrageenan and λ type carrageenan, (mixing quality ratio is κ type carrageenan: ι type carrageenan: three-dimensional porous rack λ type carrageenan=5:4:4) forming by the mass ratio of 5:1 with the chitosan of molecular weight 500,000, deacetylation 95%, porosity 95%, aperture 400～460 μ m.
A preparation method for used in tissue engineering carrageenan-chitosan composite porous support, comprises the following steps:
(1) by the carrageenan solutions of 69 ℃ of distilled water preparations 12%, the chitosan solution with PH3.4 dilute hydrochloric acid solution preparation 0.9%, is used respectively beater 8800r/min by it, making beating 30s;
(2) by above-mentioned carrageenan and chitosan solution according to carrageenan and chitosan solute mass ratio 5:1 blend, use magnetic stirring apparatus 1900r/min to stir, obtain carrageenan-chitosan complexes;
(3) by the making beating of carrageenan-chitosan complexes, add 190 μ m crystals of sodium carbonate powder (with solute mass ratio in carrageenan-chitosan complexes be 14:1), making beating again, obtains carrageenan-chitosan-carbon acid sodium complex;
(4) by 69 ℃ of water-bath 30min of carrageenan-chitosan-carbon acid sodium complex, in casting stainless steel mould, be put in freezing 4h at-40 ℃, vacuum 18Pa, 0 ℃ of lower pumping 10h lyophilization, obtain carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket;
(5) carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket is immersed in the hydrochloric acid solution of pH1.9, soak 5h, then use distilled water immersion 9h, 3h changes water once, the 18Pa 10h vacuum drying of bleeding, obtains used in tissue engineering carrageenan-chitosan composite porous support afterwards.
Embodiment 5
A kind of cartilage tissue engineered with porous support, the three-dimensional porous rack being formed by the mass ratio of 4:1 by the chitosan of κ type carrageenan and molecular weight 400,000, deacetylation 94%, porosity 96%, aperture 120～140 μ m, modulus of compressibility 0.9MPa, stretch modulus 2.1MPa.
A cartilage tissue engineered preparation method with porous support, comprises the following steps:
(1) by the carrageenan solutions of 70 ℃ of distilled water preparations 15%, with the dilute hydrochloric acid solution of PH3, prepare 1.2% chitosan solution, it is used respectively to beater 10000r/min, making beating 30s;
(2) by above-mentioned carrageenan and chitosan solution according to carrageenan and chitosan solute mass ratio 4:1 blend, use magnetic stirring apparatus 2000r/min to stir, obtain carrageenan-chitosan complexes;
(3) by the making beating of carrageenan-chitosan complexes, add 200 μ m crystals of sodium carbonate powder (with solute mass ratio in carrageenan-chitosan complexes be 15:1), making beating again, obtains carrageenan-chitosan-carbon acid sodium complex;
(4) by 70 ℃ of water-bath 30min of carrageenan-chitosan-carbon acid sodium complex, in casting stainless steel mould, be put in freezing 3h at-40 ℃, vacuum 20Pa, 0 ℃ of lower pumping 7h lyophilization, obtain carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket;
(5) carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket is immersed in the hydrochloric acid solution of pH2.5, soak 5h, then use distilled water immersion 10h, 4h changes water once, the 20Pa 7h vacuum drying of bleeding, obtains cartilage tissue engineered with carrageenan-chitosan composite porous support afterwards.
The above embodiment has only expressed several embodiment of the present invention, and it describes comparatively concrete and detailed, but can not therefore be interpreted as the restriction to the scope of the claims of the present invention.It should be pointed out that for the person of ordinary skill of the art, without departing from the inventive concept of the premise, can also make some distortion and improvement, these all belong to protection scope of the present invention.Therefore, the protection domain of patent of the present invention should be as the criterion with claims.
Claims (10)
- One kind cartilage tissue engineered with porous support, it is characterized in that: described cartilage tissue engineered with porous support by carrageenan and chitosan by (8～4): the three-dimensional porous rack that 1 mass ratio forms, porosity is 80～96%, aperture is 50～500 μ m, modulus of compressibility 0.7～1.2MPa, stretch modulus 1.0～2.2MPa.
- 2. according to claim 1 cartilage tissue engineered with porous support, it is characterized in that: described carrageenan is the mixture of κ type carrageenan κ type or κ type carrageenan, ι type carrageenan, λ type carrageenan, and mixing quality ratio is κ type carrageenan: ι type carrageenan: λ type carrageenan=(5～9): (1～5): the epoxy glue of (1～5).
- 3. according to claim 1 and 2 cartilage tissue engineered with porous support, it is characterized in that: described chitosan is the chitosan that molecular weight is 100,000～500,000, deacetylation is greater than 80%.
- 4. for the preparation of the cartilage tissue engineered method with porous support one of claim 1～3 Suo Shu, it is characterized in that: said method comprising the steps of:(1) by the carrageenan solutions of 60 ℃～70 ℃ distilled water preparations 3%～15%, with the chitosan solution of dilute acid soln preparation 0.5%～1.2%, the two is pulled an oar respectively;(2) by above-mentioned carrageenan solutions and chitosan solution blend, stir, obtain carrageenan-chitosan complexes;(3) by the making beating of carrageenan-chitosan complexes, add crystals of sodium carbonate powder, making beating again, obtains carrageenan-chitosan-carbon acid sodium complex;(4) by carrageenan-chitosan-carbon acid sodium complex water-bath 20～50min at 60 ℃～70 ℃, be cast in mould, be put in freezing 3～6h at-40 ℃～-20 ℃, lyophilization, obtains carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket;(5) carrageenan-chitosan-carbon acid sodium complex stephanoporate bracket is immersed in dilute acid soln, soak 3～5h, then use distilled water immersion 8～10h, 2～4h changes water once, and vacuum drying, obtains cartilage tissue engineered with carrageenan-chitosan composite porous support afterwards.
- 5. a kind of cartilage tissue engineered preparation method with porous support according to claim 4, is characterized in that: the carrageenan solutions that step (2) is described and chitosan solution are according to carrageenan and chitosan solute mass ratio (8～4): 1 ratio blend.
- 6. according to a kind of cartilage tissue engineered preparation method with porous support described in claim 4 or 5, it is characterized in that: in step (3), crystals of sodium carbonate powder and carrageenan-chitosan complexes solute mass ratio are (6～15): 1.
- 7. a kind of cartilage tissue engineered preparation method with porous support according to claim 6, is characterized in that: the size of the crystals of sodium carbonate powder that step (3) is described is 100～200 μ m.
- 8. a kind of cartilage tissue engineered preparation method with porous support according to claim 7, it is characterized in that: hydrochloric acid or acetic acid that the described dilute acid soln of step (1) is PH3～4, hydrochloric acid or acetic acid that the described diluted acid of step (5) is pH1.5～2.5.
- 9. a kind of cartilage tissue engineered preparation method with porous support according to claim 8, is characterized in that: the described pulping process of step (1) adopts beater, and beating rate is 8000～10000r/min, and beating time is 20～30s; Magnetic stirring apparatus is used in the described stirring of step (2), and mixing speed is 1000～2000r/min.
- 10. a kind of cartilage tissue engineered preparation method with porous support according to claim 9, is characterized in that: the described lyophilization of step (4) is vacuum 10～20Pa, 0 ℃ of lower pumping 5～10h; The described vacuum drying of step (5) is vacuum 10～20Pa 5～10h that bleeds.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108066819A (en) * | 2017-12-01 | 2018-05-25 | 浙江大学 | A kind of natural polymer hydrogel film of high intensity and preparation method thereof |
| TWI729999B (en) * | 2016-08-30 | 2021-06-11 | 艾爾生技有限公司 | Biomedical frame and method for fabricating the same |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6425918B1 (en) * | 1996-05-22 | 2002-07-30 | Ben Gurion University Of The Negev | Polysaccharide sponges for cell culture and transplantation |
| CN101322857A (en) * | 2008-07-14 | 2008-12-17 | 昆明理工大学 | Compound osseous tissue engineering stephanoporate stent material and preparation thereof |
| CN102580163A (en) * | 2012-03-21 | 2012-07-18 | 浙江大学 | Method for preparing cross-linked collagen/chitosan tissue engineering porous support in one-step freeze-drying mode |
| CN103386145A (en) * | 2013-07-31 | 2013-11-13 | 暨南大学 | Wound healing dressing containing carrageenan, and preparation method and application of wound healing dressing |
| CN103638562A (en) * | 2013-12-04 | 2014-03-19 | 上海大学 | Cartilage tissue engineering scaffold material and preparation method thereof |
-
2014
- 2014-05-07 CN CN201410190481.7A patent/CN104001213B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6425918B1 (en) * | 1996-05-22 | 2002-07-30 | Ben Gurion University Of The Negev | Polysaccharide sponges for cell culture and transplantation |
| CN101322857A (en) * | 2008-07-14 | 2008-12-17 | 昆明理工大学 | Compound osseous tissue engineering stephanoporate stent material and preparation thereof |
| CN102580163A (en) * | 2012-03-21 | 2012-07-18 | 浙江大学 | Method for preparing cross-linked collagen/chitosan tissue engineering porous support in one-step freeze-drying mode |
| CN103386145A (en) * | 2013-07-31 | 2013-11-13 | 暨南大学 | Wound healing dressing containing carrageenan, and preparation method and application of wound healing dressing |
| CN103638562A (en) * | 2013-12-04 | 2014-03-19 | 上海大学 | Cartilage tissue engineering scaffold material and preparation method thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI729999B (en) * | 2016-08-30 | 2021-06-11 | 艾爾生技有限公司 | Biomedical frame and method for fabricating the same |
| CN108066819A (en) * | 2017-12-01 | 2018-05-25 | 浙江大学 | A kind of natural polymer hydrogel film of high intensity and preparation method thereof |
| CN108066819B (en) * | 2017-12-01 | 2020-06-05 | 浙江大学 | High-strength natural polymer hydrogel film and preparation method thereof |
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