CN104004231A - Biomacromolecule interpenetrating polymer network hydrogel and preparation method thereof - Google Patents
Biomacromolecule interpenetrating polymer network hydrogel and preparation method thereof Download PDFInfo
- Publication number
- CN104004231A CN104004231A CN201410259494.5A CN201410259494A CN104004231A CN 104004231 A CN104004231 A CN 104004231A CN 201410259494 A CN201410259494 A CN 201410259494A CN 104004231 A CN104004231 A CN 104004231A
- Authority
- CN
- China
- Prior art keywords
- hydrogel
- biomacromolecule
- molecular
- network hydrogel
- network
- 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.)
- Pending
Links
Abstract
The invention discloses a biomacromolecule interpenetrating polymer network hydrogel and a preparation method of the biomacromolecule interpenetrating polymer network hydrogel. The biomacromolecule interpenetrating polymer network hydrogel is formed by crosslinking two kinds of enzymes in a catalysis mode, one is a polysaccharide macromolecule network formed by crosslinking polysaccharide macromolecules with introduced phenolic hydroxyl groups in an oxidation mode as oxidase and hydrogen peroxide catalyze phenolic hydroxyl groups, and the other is a protein or polypeptide macromolecule network formed crosslinking protein or polypeptide containing amino acid residues as transferase catalyzes peptide bonds. The two networks interpenetrate each other and form the novel interpenetrating polymer network hydrogel without bonding of chemical bonds. No chemical crosslinking agent is used in the hydrogel, and the hydrogel has excellent biocompatibility and mechanical property, can be shaped like a dry or wet film, like porous sponge or fibers, and can serve as a contact lens, a medicine release carrier, a scaffold for tissue engineering or materials for tissue repair.
Description
Technical field
The present invention is interpenetration network hydrogel being cross-linked to form by two kinds of different enzyme catalysis biomacromolecules and preparation method thereof, belongs to field of biomedical polymer materials.
Background technology
Hydrogel be a kind of can be in water swelling, keep large quantity of moisture and don't be dissolved in the three-dimensional netted polymkeric substance of water, when contacting with blood, body fluid and tissue, show good biocompatibility.Hydrogel all approaches biological tissue than other any synthesising biological material, is similar to extracellular matrix part in nature, can reduce the friction of surrounding tissue and mechanical effect after water suction, significantly improves the biology performance of material.Therefore hydrogel has great application potential aspect organizational project and regenerative medicine.
Interpenetrating(polymer)networks (IPN) hydrogel is the polymkeric substance with special performance being formed by network interpenetrating by two or more polymkeric substance.In interpenetration network hydrogel, between two component networks, there is no chemically bonded phase, each polymkeric substance all keeps characteristic separately; Meanwhile, two kinds of network interpenetratings influence each other, and have the collaborative effect that strengthens mechanical property.Therefore preparing interpenetration network hydrogel is a kind of effective ways that improve hydrogel intensity.To the research of IPN bio-medical material is existing, explore for many years both at home and abroad, conventionally adopt Chemical Crosslinking Methods to obtain IPN hydrogel.Due to chemical cross-linking agent or initiator poisonous to cell, the IPN hydrogel of preparation has potential toxicity for human body, has limited the application of interpenetration network hydrogel aspect bio-medical.
Interpenetration network hydrogel can also adopt physical crosslinking mode, as hydrophobic interaction, hydrogen bond action, ionomer etc., though do not introduce new chemical substance but be difficult to obtain degree of crosslinking uniformity, desirable, or cross-linking set may be easy to disintegrate in physiological fluid.
Glutamine transaminage (mTG) can catalytic protein or polypeptide or the molecule that contains amino-acid residue on glutamine residue with lysine residue side chain on ε-amido react, be cross-linked to form each other macromolecule hydrogel network.Horseradish peroxidase (HRP) is to take the enzyme that iron porphyrin is prothetic group, is having H
2o
2energy catalysis phenols, amine and substituent polymerization thereof while existing, on polysaccharide macro-molecular chain, exist a large amount of amino or carboxyl, can by the linked reaction of carbodiimide-mediated, introduce the structure of phenolic hydroxyl group, introduce the polysaccharide macro-molecular of phenolic hydroxyl group by the oxidation cross-linked formation macromolecule hydrogel of horseradish peroxidase enzyme catalytic phenolic hydroxyl group.
Hydrogel prepared by single enzyme-catalyzed cross-linking has been reported, but hydrogel intensity is conventionally lower, as bio-medical material, acquires a certain degree of difficulty.
Therefore, be necessary that mining inetesity is high, the interpenetration network hydrogel material of good biocompatibility.
Summary of the invention
Technical problem: residue in the cytotoxicity causing in system, the shortcoming that biocompatibility reduces in order to overcome the poor and conventional chemical cross-linking agent of hydrogel mechanical strength or initiator, the invention provides a kind of biomacromolecule interpenetration network hydrogel and application thereof, this interpenetration network hydrogel has excellent mechanical property, good biocompatibility, overcome the toxicity shortcoming that chemical cross-linking agent brings, can solve again the problem of physical crosslinking poor effect.
Technical scheme: a kind of biomacromolecule interpenetration network hydrogel, comprising:
1) polysaccharide macro-molecular that bonding is introduced phenolic hydroxyl group is by the polysaccharide macro-molecular network of phenolic hydroxyl group oxidation cross-linked formation under oxidase catalyzed effect;
2) protein or the polypeptide that contains amino-acid residue form crosslinked protein or the polypeptide network of peptide bond by transferring enzyme catalysis;
Between above-mentioned two networks, run through mutually, there is no chemical bonds, wherein the content of albumen or polypeptide network is the 0.5%-99.5% of hydrogel dry weight total mass, and the content of polysaccharide macro-molecular network is the 0.5%-99.5% of hydrogel dry weight total mass.
Described polysaccharide macro-molecular be in hyaluronic acid, dextran, chitosan, chondroitin sulfate, heparin, gum arabic, agar polysaccharide, polyfructosan, mannosans, Polygalactan, gelling gum, guar gum, Lalgine, Suleparoid, starch and derivative thereof any one or multiple.
Described bonding is introduced phenolic hydroxyl group and is specifically referred to that polysaccharide macro-molecular obtains in conjunction with any in tyrasamine, tyrosine, Dopamine HCL, homovanillic acid, gallic acid, p-hydroxyphenylaceticacid, para hydroxybenzene propionic acid by ester bond or amido linkage.
Described protein is one or more in gelatin, collagen, silk fibroin, zein, scleroproein, elastin, Saliva Orthana, glycoprotein.
Described amino-acid residue is Methionin or glutamine residue.
Described oxydase is any in peroxidase, horseradish peroxidase, tyrosine oxidase, laccase, thermolysin, phosphoesterase.
Described transferring enzyme be in glutamine transaminage, zymoplasm any.
In described hydrogel, also contain additive, this additive is one or more the mixture in cell growth factor, bone morphogenetic protein, VITAMIN, Regular Insulin, steroid hormone, dexamethasone, medicine or inorganic salt, and the content of additive is the 0.01%-3% of hydrogel total mass.
A method of preparing described biomacromolecule interpenetration network hydrogel, step is:
1) polysaccharide macro-molecular that preparation contains phenolic hydroxyl group; Concrete method for making is shown in embodiment 1～4;
2) polypeptide of take the polysaccharide macro-molecular and the protein that contain phenolic hydroxyl group or containing amino-acid residue is prepared biomacromolecule interpenetration network hydrogel as reaction raw materials under the catalysis of oxydase, transferring enzyme and hydrogen peroxide.Wherein, the polysaccharide macro-molecular that bonding is introduced phenolic hydroxyl group is by phenolic hydroxyl group oxidation cross-linked formation polysaccharide macro-molecular network under oxidase catalyzed effect; Protein or the polypeptide that contains amino-acid residue form crosslinked protein or the polypeptide network of peptide bond by transferring enzyme catalysis.
Beneficial effect:
1, the present invention on polysaccharide macro-molecular chain, introduce phenolic hydroxyl group by oxidase catalyzed phenolic hydroxyl group oxidation cross-linked and albumen or contain Methionin and the polypeptide of glutamine residue forms interpenetration network hydrogel by transferring enzyme catalysis peptide bond is crosslinked, two network cooperating effects have improved the mechanical strength of hydrogel.
3, the present invention's starting material used are biomacromolecule or the polypeptide that come from nature, and the interpenetration network hydrogel obtaining has excellent biocompatibility.
4, the method that the present invention adopts two kinds of enzyme-catalyzed cross-linkings to prepare interpenetration network hydrogel had both overcome the toxicity that chemical cross-linking agent brings, and had solved again the shortcoming of physical crosslinking agent poor effect.
5, interpenetration network hydrogel of the present invention can be processed into dry state or hygrometric state is membranaceous, porous spongy, fibrous, can be used as tissue engineering bracket or tissue renovation material.
6, interpenetration network hydrogel of the present invention can contain cell growth factor, VITAMIN, steroid hormone, medicine or inorganic salt one or more as additive, give this hydrogel various Biomedical functions.
7, biomacromolecule interpenetration network hydrogel of the present invention, as contact lense, has excellent anti-dewatering and comfortable wearing.
Accompanying drawing explanation
Fig. 1 is that polysaccharide macro-molecular is introduced the structure diagram after phenolic hydroxyl group.
Fig. 2 is the enzyme-catalyzed cross-linking mechanism sketch of introducing the polysaccharide macro-molecular of phenolic hydroxyl group.
Fig. 3 is the enzyme-catalyzed cross-linking mechanism sketch of protein or polypeptide macromolecule.
Fig. 4 introduces the polysaccharide macro-molecular of phenolic hydroxyl group and the sketch of albumen or polypeptide macromolecule formation interpenetration network hydrogel.
Fig. 5 is chitosan-para hydroxybenzene propionic acid of embodiment 1 preparation
1h NMR collection of illustrative plates.
Embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is done further and explained.The following example is only for the present invention is described, but is not used for limiting practical range of the present invention.
Biomacromolecule interpenetration network hydrogel of the present invention is the biomacromolecule interpenetration network hydrogel that adopts two kinds of different enzyme-catalyzed cross-linkings to form, this interpenetration network hydrogel has good biocompatibility and mechanical strength, can be used for cell, tissue culture, as tissue engineering bracket or tissue renovation material, can make membranaceous, three-dimensional porous shape, or fibrous etc., can be used as contact lense.
A kind of biomacromolecule interpenetration network hydrogel of the present invention, this interpenetration network hydrogel comprises: the polysaccharide macro-molecular network that 1) polysaccharide macro-molecular of bonding introducing phenolic hydroxyl group is cross-linked to form under oxidase catalyzed effect by phenolic hydroxyl group; 2) protein or contain Methionin and protein or polypeptide macromolecule network that the polypeptide of glutamine residue forms by transferring enzyme catalytic crosslinking; Between above-mentioned two networks, run through mutually, there is no chemical bonds, wherein the content of albumen or polypeptide or synthetic macromolecule network is the 0.5%-99.5% of hydrogel dry weight total mass, and the content of polysaccharide macro-molecular network is the 0.5%-99.5% of hydrogel dry weight total mass.
Described polysaccharide is one or more in hyaluronic acid, dextran, chitosan, chondroitin sulfate, heparin, gum arabic, agar polysaccharide, polyfructosan, mannosans, Polygalactan, gelling gum, guar gum, Lalgine, Suleparoid, starch and derivative thereof.
The phenolic hydroxyl structure of introducing on polysaccharide macro-molecular chain can be a kind of by the tyrasamine of ester bond or amido linkage combination, tyrosine, Dopamine HCL, homovanillic acid, gallic acid, p-hydroxyphenylaceticacid, para hydroxybenzene propionic acid.
Described albumen is one or more in gelatin, collagen, silk fibroin, zein, scleroproein, elastin, Saliva Orthana, glycoprotein.
Described oxydase is peroxidase, horseradish peroxidase, tyrosine oxidase, laccase, thermolysin, phosphoesterase.Described transferring enzyme is that glutamine transaminage (TG), glutamine turn ammonia isozyme, zymoplasm.Glutamine transaminage can come from tissue, can be also microorganism fermentation preparation, is designated as mTG.
A kind of biomacromolecule interpenetration network hydrogel of the present invention also contains additive, this additive is one or more mixture of cell growth factor, bone morphogenetic protein, VITAMIN, Regular Insulin, steroid hormone, dexamethasone, medicine or inorganic salt, and the content of additive is the 0.01%-3% of hydrogel total mass.
A kind of biomacromolecule interpenetration network hydrogel of the present invention can be membranaceous, fibrous, the porous spongy of hygrometric state or dry state, as contact lens, drug release carrier, tissue engineering bracket or tissue renovation material.
A method of preparing described biomacromolecule interpenetration network hydrogel, step is:
1) polysaccharide macro-molecular that preparation contains phenolic hydroxyl group; Concrete method for making is shown in embodiment 1～4;
2) polypeptide of take the polysaccharide macro-molecular and the protein that contain phenolic hydroxyl group or containing amino-acid residue is prepared biomacromolecule interpenetration network hydrogel as reaction raw materials under the catalysis of oxydase, transferring enzyme and hydrogen peroxide; Bonding is introduced the polysaccharide macro-molecular of phenolic hydroxyl group by phenolic hydroxyl group oxidation cross-linked formation polysaccharide macro-molecular network under oxidase catalyzed effect; Protein or the polypeptide that contains amino-acid residue form crosslinked protein or the polypeptide network of peptide bond by transferring enzyme catalysis.
Embodiment 1
The preparation of chitosan-para hydroxybenzene propionic acid (CHPA)
Take water-soluble chitosan (0.88g, 5mmol) and be dissolved in 150ml deionized water, with magnetic stirring apparatus constant temperature, be stirred to completely and dissolve.Take para hydroxybenzene propionic acid (1.66g, 10mmol), 1-ethyl-(3-dimethylaminopropyl) phosphinylidyne diimmonium salt hydrochlorate (EDCHCl) (2.73g, 15mmol), N-hydroxy-succinamide (NHS) (1.72g, 15mmol) be dissolved in DMF (DMF) aqueous solution of 100ml, wherein DMF: water (3:2, V:V), room temperature activation 1h (pH=4.7), joins activation solution in chitosan solution stirred overnight at room temperature.In reaction solution immigration dialysis tubing, distill water dialysis is 3 days, and dialyzate adjusts pH to separate out product, and vacuum-drying obtains the chitosan of product bonding para hydroxybenzene propionic acid, is below designated as chitosan-para hydroxybenzene propionic acid 0.7g, productive rate 80%.Warp
1the substitution value that H NMR measures para hydroxybenzene propionic acid in product is 15%, as shown in Figure 5.
Embodiment 2
The preparation of hyaluronic acid-tyrasamine (HA-Tyr)
Take hyaluronic acid (2g, 5mmol) and be dissolved in 100ml deionized water, with magnetic stirring apparatus constant temperature, be stirred to completely and dissolve.Take tyrasamine (1.68g, 10mmol), EDCHCl (2.73g, 15mmol), NHS (1.72g, 15mmol) is dissolved in the distilled water of 100ml, room temperature activation 1h (pH=4.7), joins activation solution in hyaluronic acid solution stirred overnight at room temperature.In reaction solution immigration dialysis tubing, distill water dialysis is 3 days, and dialyzate adjusts pH to separate out product, and vacuum-drying obtains the hyaluronic acid of product bonding tyrasamine, is below designated as hyaluronic acid-tyrasamine.
Embodiment 3
The preparation of heparin-tyrasamine (Hep-Tyr)
Take heparin (0.5g) and be dissolved in 100ml deionized water, with magnetic stirring apparatus constant temperature, be stirred to completely and dissolve.Take tyrasamine (1.68g, 10mmol), EDCHCl (2.73g, 15mmol), NHS (1.72g, 15mmol) is dissolved in the distilled water of 100ml, room temperature activation 1h (pH=4.7), joins activation solution in heparin solution stirred overnight at room temperature.In reaction solution immigration dialysis tubing, distill water dialysis is 3 days, and dialyzate adjusts pH to separate out product, and vacuum-drying obtains the heparin of product bonding tyrasamine, is below designated as heparin-tyrasamine.
Embodiment 4
The preparation of Lalgine-Dopamine HCL (Alg-Dopa)
Take Lalgine (2g, 5mmol) and be dissolved in 200ml deionized water, with magnetic stirring apparatus constant temperature, be stirred to completely and dissolve.Claim Dopamine HCL (2g, 10mmol), EDCHCl (2.73g, 15mmol), NHS (1.72g, 15mmol) is dissolved in the distilled water of 100ml, and room temperature activation 1h (pH=4.7), joins activation solution in Lalgine solution stirred overnight at room temperature.In reaction solution immigration dialysis tubing, distill water dialysis is 3 days, and dialyzate adjusts pH to separate out product, and vacuum-drying obtains the Lalgine of product bonding Dopamine HCL, is below designated as Lalgine-Dopamine HCL.
Embodiment 5
Take in acetic acid-sodium-acetate buffer (pH=5.4) that 0.1g chitosan-para hydroxybenzene propionic acid (CHPA) is dissolved in 10ml and obtain 1% the CHPA aqueous solution, taking 1g gelatin joins in the above-mentioned CHPA aqueous solution, 40 ℃ of heating in water bath dissolve, standing froth breaking, the CHPA being uniformly mixed and aqueous gelatin solution.
Adding 0.5ml concentration is the glutamine transaminage of 100U/ml, the horseradish peroxidase that 0.1ml concentration is 500U/ml, and adding 0.06ml concentration is the hydrogen peroxide (H of 25mM
2o
2), Quick uniform mixes, and is placed in columnar mould, and cross-linking reaction 40min at 40 ℃, obtains membranaceous biomacromolecule interpenetration network hydrogel.
Repeatedly wash this hydrogel, remove residual H
2o
2, after acetic acid, sodium-acetate, be kept in distilled water, obtain gelatin/CHPA interpenetration network hydrogel of two kinds of different enzyme-catalyzed cross-linkings.Gelatin is cross-linked and is obtained protein macromolecule network by glutamine transaminage, and CHPA is oxidized the crosslinked polysaccharide macro-molecular network that obtains of phenolic hydroxyl group by horseradish peroxidase.
Embodiment 6
Take in acetic acid-sodium-acetate buffer (pH=5.4) that 0.1g Lalgine-Dopamine HCL (Alg-Dopa) is dissolved in 10ml and obtain 1% the Alg-Dopa aqueous solution, taking 1g gelatin joins in the above-mentioned Alg-Dopa aqueous solution, 40 ℃ of heating in water bath dissolve, standing froth breaking, the Alg-Dopa being uniformly mixed and aqueous gelatin solution.
Adding 0.5ml concentration is the glutamine transaminage of 100U/ml, the horseradish peroxidase that 0.1ml concentration is 500U/ml, and adding 0.06ml concentration is the H of 25mM
2o
2, Quick uniform mixes, and in columnar mould, cross-linking reaction 40min at 40 ℃, obtains membranaceous biomacromolecule interpenetration network hydrogel.
Repeatedly wash this hydrogel, remove residual H
2o
2, after acetic acid, sodium-acetate, be kept in distilled water, obtain gelatin/Alg-Dopa interpenetration network hydrogel of two kinds of different enzyme-catalyzed cross-linkings.Gelatin is cross-linked and is obtained protein macromolecule network by glutamine transaminage, and Alg-Dopa is cross-linked and is obtained polysaccharide macro-molecular network by horseradish peroxidase.
Embodiment 7
Take in acetic acid-sodium-acetate buffer (pH=5.4) that 0.1g hyaluronic acid-tyrasamine (HA-Tyr) is dissolved in 10ml and obtain 1% the HA-Tyr aqueous solution, taking 1g gelatin joins in the above-mentioned HA-Tyr aqueous solution, 40 ℃ of heating in water bath dissolve, standing froth breaking, the HA-Tyr being uniformly mixed and aqueous gelatin solution.
Adding 0.5ml concentration is the glutamine transaminage of 100U/ml, the horseradish peroxidase that 0.1ml concentration is 500U/ml, and adding 0.06ml concentration is the H of 25mM
2o
2, Quick uniform mixes, and in columnar mould, cross-linking reaction 40min at 40 ℃, obtains membranaceous biomacromolecule interpenetration network hydrogel.
Repeatedly wash this hydrogel, remove residual H
2o
2, after acetic acid, sodium-acetate, be kept in distilled water, obtain gelatin/HA-Tyr interpenetration network hydrogel of two kinds of different enzyme-catalyzed cross-linkings.Gelatin is cross-linked and is obtained protein macromolecule network by glutamine transaminage, and HA-Tyr is oxidized the crosslinked polysaccharide macro-molecular network that obtains of phenolic hydroxyl group by horseradish peroxidase.
Embodiment 8
Take in acetic acid-sodium-acetate buffer (pH=5.4) that 0.1g heparin-tyrasamine (Hep-Tyr) is dissolved in 10ml and obtain 1% the Hep-Tyr aqueous solution, taking 1g gelatin joins in the above-mentioned Hep-Tyr aqueous solution, 40 ℃ of heating in water bath dissolve, standing froth breaking, the Hep-Tyr being uniformly mixed and aqueous gelatin solution.
Adding 0.5ml concentration is the glutamine transaminage of 100U/ml, the horseradish peroxidase that 0.1ml concentration is 500U/ml, and adding 0.06ml concentration is the H of 25mM
2o
2, Quick uniform mixes, and in columnar mould, cross-linking reaction 40min at 40 ℃, obtains column biomacromolecule interpenetration network hydrogel.
Repeatedly wash this hydrogel, remove residual H
2o
2, after acetic acid, sodium-acetate, be kept in distilled water, obtain gelatin/Hep-Tyr interpenetration network hydrogel of two kinds of different enzyme-catalyzed cross-linkings.Gelatin is cross-linked and is obtained protein macromolecule network by glutamine transaminage, and Hep-Tyr is cross-linked and is obtained polysaccharide macro-molecular network by horseradish peroxidase.
Embodiment 9
Take in acetic acid-sodium-acetate buffer (pH=5.4) that 0.1g chitosan-para hydroxybenzene propionic acid (CHPA) is dissolved in 10ml and obtain 1% the CHPA aqueous solution, taking 1g zein joins in the above-mentioned CHPA aqueous solution, 40 ℃ of heating in water bath dissolve, standing froth breaking, the CHPA being uniformly mixed and the zein aqueous solution.
Adding 0.5ml concentration is the glutamine transaminage of 100U/ml, the horseradish peroxidase that 0.1ml concentration is 500U/ml, and adding 0.06ml concentration is the H of 25mM
2o
2, Quick uniform mixes, and in mould, cross-linking reaction 40min at 40 ℃, obtains biomacromolecule interpenetration network hydrogel.
Repeatedly wash this hydrogel, remove residual H
2o
2, after acetic acid, sodium-acetate, be kept in distilled water, obtain zein/CHPA interpenetration network hydrogel of two kinds of different enzyme-catalyzed cross-linkings.Zein is cross-linked and is obtained protein macromolecule network by glutamine transaminage, and CHPA is oxidized the crosslinked polysaccharide macro-molecular network that obtains of phenolic hydroxyl group by horseradish peroxidase.
Embodiment 10
Take in acetic acid-sodium-acetate buffer (pH=5.4) that 0.1g chitosan-para hydroxybenzene propionic acid (CHPA) is dissolved in 10ml and obtain the CHPA aqueous solution, take that 1g contains Methionin and glutamine small-molecular peptides joins in the above-mentioned CHPA aqueous solution, 40 ℃ of heating in water bath dissolve, standing froth breaking.Adding 0.5ml concentration is the glutamine transaminage of 50U/ml, the horseradish peroxidase that 0.1ml concentration is 500U/ml, and Quick uniform mixes, and in mould, cross-linking reaction 40min at 40 ℃, obtains biomacromolecule interpenetration network hydrogel.
Repeatedly wash this hydrogel, remove after residual acetic acid, sodium-acetate, be kept in distilled water, obtain polypeptide/CHPA interpenetration network hydrogel of two kinds of different enzyme-catalyzed cross-linkings, small-molecular peptides is cross-linked and is obtained polypeptide macromolecule network by glutamine transaminage, and CHPA is oxidized the crosslinked polysaccharide macro-molecular network that obtains of phenolic hydroxyl group by horseradish peroxidase.
Embodiment 11
Take 0.1g hyaluronic acid-tyrasamine (HA-Tyr) and be dissolved in the HA-Tyr aqueous solution that obtains 1% in 10m distilled water, taking 1g gelatin joins in the above-mentioned HA-Tyr aqueous solution, 40 ℃ of heating in water bath dissolve, standing froth breaking, the HA-Tyr being uniformly mixed and aqueous gelatin solution.
Adding 0.5ml concentration is the glutamine transaminage of 100U/ml, the tyrosine oxidase that 0.1ml concentration is 500U/ml, and adding 0.06ml concentration is the H of 25mM
2o
2, Quick uniform mixes, and in mould, cross-linking reaction 40min at 40 ℃, obtains biomacromolecule interpenetration network hydrogel.
Repeatedly wash this hydrogel, remove residual H
2o
2after, be kept in distilled water, gelatin/HA-Tyr the interpenetration network hydrogel that obtains two kinds of different enzyme-catalyzed cross-linkings, gelatin is cross-linked and is obtained protein macromolecule network by glutamine transaminage, and HA-Tyr is cross-linked and is obtained polysaccharide macro-molecular network by horseradish peroxidase.
Embodiment 12
Take in acetic acid-sodium-acetate buffer (pH=5.4) that 0.1g chitosan-para hydroxybenzene propionic acid (CHPA) is dissolved in 10ml and obtain the CHPA aqueous solution, taking 1g gelatin joins in the above-mentioned CHPA aqueous solution, 40 ℃ of heating in water bath dissolve, standing froth breaking, the CHPA being uniformly mixed and aqueous gelatin solution.
Adding 0.25ml concentration is the zymoplasm of 200U/ml, the horseradish peroxidase that 0.1ml concentration is 500U/ml, and adding 0.06ml concentration is the H of 25mM
2o
2, Quick uniform mixes, and in mould, cross-linking reaction 40min at 40 ℃, obtains biomacromolecule interpenetration network hydrogel.
Repeatedly wash this hydrogel, remove residual H
2o
2, after acetic acid, sodium-acetate, be kept in distilled water, gelatin/CHPA the interpenetration network hydrogel that obtains two kinds of different enzyme-catalyzed cross-linkings, gelatin is cross-linked and is obtained protein macromolecule network by zymoplasm, and CHPA is cross-linked and is obtained polysaccharide macro-molecular network by horseradish peroxidase.
Embodiment 13
Take in acetic acid-sodium-acetate buffer (pH=5.4) that 0.1g chitosan-para hydroxybenzene propionic acid (CHPA) is dissolved in 10ml and obtain the CHPA aqueous solution, taking 1g gelatin joins in the above-mentioned CHPA aqueous solution, 40 ℃ of heating in water bath dissolve, standing froth breaking, the CHPA being uniformly mixed and aqueous gelatin solution.
Adding 0.5ml concentration is the glutamine transaminage of 100U/ml, the laccase that 0.1ml concentration is 500U/ml, and Quick uniform mixes, and in mould, cross-linking reaction 40min at 40 ℃, obtains biomacromolecule interpenetration network hydrogel.
Repeatedly wash this hydrogel, remove after residual acetic acid, sodium-acetate, be kept in distilled water, obtain gelatin/CHPA interpenetration network hydrogel of two kinds of different enzyme-catalyzed cross-linkings, gelatin is cross-linked and is obtained protein macromolecule network by glutamine transaminage, and CHPA is cross-linked and is obtained polysaccharide macro-molecular network by laccase.
Embodiment 14
Take in acetic acid-sodium-acetate buffer (PH=5.4) that 0.2g chitosan-para hydroxybenzene propionic acid (CHPA) is dissolved in 20ml and obtain the CHPA aqueous solution, taking 2g gelatin joins in the above-mentioned CHPA aqueous solution, 40 ℃ of heating in water bath dissolve, after standing froth breaking, adding 0.2ml concentration is the horseradish peroxidase Quick uniform mixing of 500U/ml, the CHPA being uniformly mixed and aqueous gelatin solution.
40 ℃ by above-mentioned mixed solution with micro-injection pump take 50ml/h speed injection to containing glutamine transaminage concentration be 15U/ml, H
2o
2concentration is in the aqueous solution of 0.8mM, and cross-linking reaction 40min at 40 ℃, obtains fibrous interpenetration network hydrogel.Repeatedly wash this hydrogel, remove residual H
2o
2, after acetic acid, sodium-acetate, be kept in distilled water, obtain gelatin/CHPA interpenetration network hydrogel fiber of two kinds of different enzyme-catalyzed cross-linkings.
Embodiment 15
Take in acetic acid-sodium-acetate buffer (PH=5.4) that 0.2g chitosan-para hydroxybenzene propionic acid (CHPA) is dissolved in 20ml and obtain 1% the CHPA aqueous solution, taking 2g gelatin joins in the above-mentioned CHPA aqueous solution, after 40 ℃ of heating in water bath dissolve, separately add 2000IU recombinant human epidermal growth factor, stir, standing froth breaking, obtains the CHPA and the aqueous gelatin solution that contain recombinant human epidermal growth factor.
Adding 1ml concentration is the glutamine transaminage of 100U/ml, the horseradish peroxidase that 0.2ml concentration is 500U/ml, and adding 0.12ml concentration is the H of 25mM
2o
2, Quick uniform mixes, and in column grinding tool, cross-linking reaction 40min at 40 ℃, obtains membranaceous interpenetration network hydrogel.Repeatedly wash this hydrogel, remove residual H
2o
2, after acetic acid, sodium-acetate, be kept in distilled water, obtain gelatin/CHPA interpenetration network hydrogel of two kinds of different enzyme-catalyzed cross-linkings.By above-mentioned hydrogel, further through gas foaming, vacuum lyophilization, obtains gelatin/CHPA interpenetrating(polymer)networks porous material, can be used as drug delivery system material.
Embodiment 16
Take in acetic acid-sodium-acetate buffer (PH=5.4) that 0.1g chitosan-para hydroxybenzene propionic acid (CHPA) is dissolved in 10ml and obtain the CHPA aqueous solution, adding 0.5ml concentration is that the glutamine transaminage of 100U/ml evenly mixes.Take 1g gelatin and join in the above-mentioned CHPA aqueous solution, 40 ℃ of heating in water bath dissolve, and after standing froth breaking, adding 0.2ml concentration is the horseradish peroxidase of 500U/ml and the H that 0.12ml concentration is 8mM
2o
2evenly mix.After above-mentioned two kinds of mixed solution Quick uniforms mix, inject contact lens mould, cross-linking reaction 40min at 40 ℃, obtains interpenetration network hydrogel contact lens.Repeatedly wash this hydrogel contact glass, remove residual H
2o
2, after acetic acid, sodium-acetate, be kept in distilled water, obtain gelatin/CHPA interpenetration network hydrogel contact lens of two kinds of different enzyme-catalyzed cross-linkings.
Embodiment 17
Take in acetic acid-sodium-acetate buffer (pH=5.4) that 0.2g chitosan-para hydroxybenzene propionic acid (CHPA) is dissolved in 20ml and obtain 1% the CHPA aqueous solution, taking 2g gelatin joins in the above-mentioned CHPA aqueous solution, 40 ℃ of heating in water bath dissolve, standing froth breaking, the CHPA being uniformly mixed and aqueous gelatin solution.
Adding 1ml concentration is the glutamine transaminage of 100U/ml, the horseradish peroxidase that 0.2ml concentration is 500U/ml, and adding 0.12ml concentration is the H of 25mM
2o
2, Quick uniform mixes, and in column grinding tool, cross-linking reaction 40min at 40 ℃, obtains membranaceous interpenetration network hydrogel.Repeatedly wash this hydrogel, remove residual H
2o
2, after acetic acid, sodium-acetate, further through liquid-nitrogen freeze drying, obtain gelatin/CHPA porous spongy biomacromolecule interpenetration network hydrogel of two kinds of different enzyme-catalyzed cross-linkings, can be used as tissue engineering bracket, tissue renovation material.
Embodiment 18
Preparing gelatin concentration is 10%, and chitosan-para hydroxybenzene propionic acid (CHPA) content is respectively 0%, 0.5%, and gelatin/CHPA mixing solutions of 1%, 1.5%, adds TG enzyme, HRP enzyme, H
2o
2make final concentration be respectively 10U/mL, 5U/mL, 0.15mM.By the composition in table 1, be mixed to join in mould, cross-linking reaction 40min at 40 ℃, obtains the interpenetration network hydrogel of two kinds of enzyme-catalyzed cross-linkings.
The composition of table 1. gelatin/CHPA interpenetration network hydrogel
| Sample | Gelatin (mg/mL) | Chitosan-para hydroxybenzene propionic acid (mg/mL) |
| Gelatin | 100 | 0 |
| G-C1 | 100 | 5 |
| G-C2 | 100 | 10 |
| G-C3 | 100 | 15 |
Compression and the stretching mechanical test result of gelatin/CHPA interpenetration network hydrogel show, the mechanical property of interpenetration network hydrogel significantly strengthens, and the increase along with CHPA content, stretching and compressive strength all increase, the tensile strength of gelatin hydrogel and compressive strength are respectively 0.189MPa and 2.05MPa, and the tensile strength of G-C3 interpenetration network hydrogel and compressive strength are respectively 0.66MPa and 4.01MPa, be respectively 3.5 times and 2 times of gelatin hydrogel, elongation at break is up to 211%.
The crosslinked crosslinked crosslinked gelatin/CHPA interpenetration network hydrogel of CHPA hydrogel, mTG/HRP of gelatin hydrogel, HRP enzyme of mTG enzyme is placed in PBS solution and soaks 24h, CHPA hydrogel is swollen rotten shape, and gelatin/CHPA interpenetration network hydrogel tensile strength is without obvious decline.
Embodiment 19
Gelatin/CHPA hydrogel cross-linking process carries out rheology test (forming as table 1).Gelatin under the effect of mTG enzyme 40 ℃ crosslinked, in 25min, 10% gelatin elastic modulus G ' is increased to 1116Pa from 0.01Pa; The gelatin glutamine residue on its molecular chain and lysine residue covalent cross-linking under the effect of mTG enzyme are described, have formed crosslinked network.
10% gelatin solution containing 0.5%CHPA is crosslinked at 40 ℃.The gelatin viscous modulus that contains 0.5%CHPA is 3 times (25.4Pa vs9.24Pa) of pure gelatin, shows that the increase of CHPA content causes the viscous modulus of hydrogel to enlarge markedly, and has obviously changed its SOLUTION PROPERTIES.After 25min similar (1116Pa, the 1032Pa) of the elastic modulus G ' of gel and pure gelatin, show that CHPA does not affect the formation of gelatin cross-linked network, do not belong to the constituent of gelatin network, CHPA keeps solution state in gelatin network.This hydrogel network is called semi-intercrossing network.
Respectively containing 10% gelatin solution of 0.5%, 1%, 1.5%CHPA and mTG/HRP enzyme, H
2o
2at 40 ℃, temperature is bathed.In 25min, elastic modulus G ' obviously increases (1675Pa, 2847Pa and 3372Pa), viscous modulus G along with the increase of CHPA concentration " along with the also obviously increase (28.68Pa, 66.23Pa and 143.8Pa) of increase of CHPA concentration, show that HRP enzyme is at H
2o
2while existing, participation system is crosslinked, has obviously improved elastic property and the adhesion properties of hydrogel.Obtained gelatin/CHPA interpenetration network hydrogel.
Embodiment 20
Gelatin/CHPA interpenetration network hydrogel vitro enzyme Study on degradation
Hydrogel (forming as table 1) is made cylindric sample freeze-drying and is weighed, and its quality is designated as W
0, put into culture dish, add the PBS5ml containing papoid, 37 ℃, the vibration of 80rpm shaking table, respectively at 2h, 4h, 6h, 8h, 10h time point takes out some samples, distilled water flushing, freeze-drying is weighed, and its quality is designated as W
t.Every group of 3 parallel sampleses, averaged.Calculate remaining rate.Result shows, along with the increase degradation rate of CHPA concentration reduces, shows that the anti-degradation property of the interpenetration network hydrogel of two kinds of enzyme-catalyzed cross-linkings is better than the gelatin semi-interpenetrating network aqueous gel of single enzyme-catalyzed cross-linking.
Embodiment 21
Gelatin/CHPA interpenetration network hydrogel biological evaluation
Cell toxicity test: spend the night with the lixiviate of DMEM substratum after hydrogel (forming as table 1) ultraviolet sterilization 2h, vat liquor is cultivated L929 cell, and mtt assay is measured the relative appreciation rate of cell.Result demonstration, the relative appreciation rate of cell of the 1st day is all more than 85%, and the cell appreciation rate of the 3rd, 5 days, all more than 95%, shows that gelatin/CHPA interpenetration network hydrogel has good biocompatibility.
Cell adhesion test: hydrogel ultraviolet sterilization 2h, soaks 4h with substratum.Inoculation 10
5the L929 cell of/ml on aquagel membrane, at 37 ℃, 5%CO
2in incubator, cultivate 5 days.Result demonstration, cell is fusiformis and aquagel membrane surface tight adhesion, along with incubation time increases, the continuous division growth of cell, migration, in the time of the 5th day, cell almost covers with aquagel membrane surface, and mutual adjacent formation is thickly dotted netted.
Aquagel membrane by culturing cell after 5 days takes out, and glutaraldehyde is fixed, scanning electron microscopic observation after freeze-drying.Result shows, cell is single shuttle shape or bulk tight adhesion in gel film surface, and visible growth in order.Biological evaluation result shows that glue/CHPA interpenetration network hydrogel has excellent cellular affinity.
Embodiment 22
Take in acetic acid-sodium-acetate buffer (pH=5.4) that 0.1g heparin-tyrasamine (Hep-Tyr) is dissolved in 10ml and obtain 1% the Hep-Tyr aqueous solution, taking 1g gelatin joins in the above-mentioned Hep-Tyr aqueous solution, 40 ℃ of heating in water bath dissolve, standing froth breaking, the Hep-Tyr being uniformly mixed and aqueous gelatin solution.
Adding 0.5ml concentration is the glutamine transaminage of 100U/ml, the horseradish peroxidase that 0.1ml concentration is 500U/ml, and adding 0.06ml concentration is the H of 25mM
2o
2, Quick uniform mixes, and in columnar mould, cross-linking reaction 40min at 40 ℃, obtains column biomacromolecule interpenetration network hydrogel.
Repeatedly wash this hydrogel, remove residual H
2o
2, after acetic acid, sodium-acetate, be immersed in the solution that contains cell growth factor, obtain the gelatin/Hep-Tyr interpenetration network hydrogel that contains somatomedin of two kinds of different enzyme-catalyzed cross-linkings.
Claims (10)
1. a biomacromolecule interpenetration network hydrogel, is characterized in that, comprising:
1) polysaccharide macro-molecular that bonding is introduced phenolic hydroxyl group is by the polysaccharide macro-molecular network of phenolic hydroxyl group oxidation cross-linked formation under oxidase catalyzed effect;
2) protein or the polypeptide that contains amino-acid residue form crosslinked protein or the polypeptide network of peptide bond by transferring enzyme catalysis;
Between above-mentioned two networks, run through mutually, there is no chemical bonds, wherein the content of albumen or polypeptide network is the 0.5%-99.5% of hydrogel dry weight total mass, and the content of polysaccharide macro-molecular network is the 0.5%-99.5% of hydrogel dry weight total mass.
2. a kind of biomacromolecule interpenetration network hydrogel according to claim 1, it is characterized in that, described polysaccharide macro-molecular be in hyaluronic acid, dextran, chitosan, chondroitin sulfate, heparin, gum arabic, agar polysaccharide, polyfructosan, mannosans, Polygalactan, gelling gum, guar gum, Lalgine, Suleparoid, starch and derivative thereof any one or multiple.
3. a kind of biomacromolecule interpenetration network hydrogel according to claim 1, it is characterized in that, described bonding is introduced phenolic hydroxyl group and is specifically referred to that polysaccharide macro-molecular obtains in conjunction with any in tyrasamine, tyrosine, Dopamine HCL, homovanillic acid, gallic acid, p-hydroxyphenylaceticacid, para hydroxybenzene propionic acid by ester bond or amido linkage.
4. a kind of biomacromolecule interpenetration network hydrogel according to claim 1, is characterized in that, described protein is one or more in gelatin, collagen, silk fibroin, zein, scleroproein, elastin, Saliva Orthana, glycoprotein.
5. a kind of biomacromolecule interpenetration network hydrogel according to claim 1, is characterized in that, described amino-acid residue is Methionin or glutamine residue.
6. a kind of biomacromolecule interpenetration network hydrogel according to claim 1, is characterized in that, described oxydase is any in peroxidase, horseradish peroxidase, tyrosine oxidase, laccase, thermolysin, phosphoesterase.
7. a kind of biomacromolecule interpenetration network hydrogel according to claim 1, is characterized in that, described transferring enzyme be in glutamine transaminage, zymoplasm any.
8. a kind of biomacromolecule interpenetration network hydrogel according to claim 1, it is characterized in that, in described hydrogel, also contain additive, this additive is one or more the mixture in cell growth factor, bone morphogenetic protein, VITAMIN, Regular Insulin, steroid hormone, dexamethasone, medicine or inorganic salt, and the content of additive is the 0.01%-3% of hydrogel total mass.
9. the arbitrary described a kind of biomacromolecule interpenetration network hydrogel of claim 1～8 is with membranaceous, fibrous, the porous spongy of hygrometric state or dry state, for contact lens, drug release carrier, tissue engineering bracket or tissue renovation material.
10. a method of preparing the arbitrary described biomacromolecule interpenetration network hydrogel of claim 1～8, is characterized in that, step is:
1) polysaccharide macro-molecular that preparation contains phenolic hydroxyl group;
2) polypeptide of take the polysaccharide macro-molecular and the protein that contain phenolic hydroxyl group or containing amino-acid residue is prepared biomacromolecule interpenetration network hydrogel as reaction raw materials under the catalysis of oxydase, transferring enzyme and hydrogen peroxide; Bonding is introduced the polysaccharide macro-molecular of phenolic hydroxyl group by phenolic hydroxyl group oxidation cross-linked formation polysaccharide macro-molecular network under oxidase catalyzed effect; Protein or the polypeptide that contains amino-acid residue form crosslinked protein or the polypeptide network of peptide bond by transferring enzyme catalysis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410259494.5A CN104004231A (en) | 2014-06-12 | 2014-06-12 | Biomacromolecule interpenetrating polymer network hydrogel and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410259494.5A CN104004231A (en) | 2014-06-12 | 2014-06-12 | Biomacromolecule interpenetrating polymer network hydrogel and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104004231A true CN104004231A (en) | 2014-08-27 |
Family
ID=51365151
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410259494.5A Pending CN104004231A (en) | 2014-06-12 | 2014-06-12 | Biomacromolecule interpenetrating polymer network hydrogel and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104004231A (en) |
Cited By (51)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104667349A (en) * | 2015-02-06 | 2015-06-03 | 福州大学 | Growth factor-loading silk fibroin/collagen bracket material and preparation method thereof |
| CN105131632A (en) * | 2015-09-21 | 2015-12-09 | 高瑞杰 | Environment-friendly food package film based on modified elastic aloe gel micro powder and preparation method |
| CN105254906A (en) * | 2015-11-13 | 2016-01-20 | 东南大学 | Method for preparing chemical crosslinking and metal ion complexing type interpenetrating network hydrogel |
| CN105802251A (en) * | 2016-04-25 | 2016-07-27 | 成都大学 | Self-assembly collagen template tissue engineering material as well as preparation method and application thereof |
| CN105944150A (en) * | 2016-03-25 | 2016-09-21 | 华东理工大学 | Hydrogel, artificial skin manufactured through application of hydrogel and manufacturing method of artificial skin |
| CN106179272A (en) * | 2016-08-10 | 2016-12-07 | 合众（佛山）化工有限公司 | A kind of novel porous nano carbon microsphere composite aquogel and preparation method thereof |
| CN106700099A (en) * | 2016-11-23 | 2017-05-24 | 华中科技大学同济医学院附属协和医院 | Colorless and transparent silk sericin hydrogel, stent prepared therefrom and application thereof |
| CN106750393A (en) * | 2016-11-15 | 2017-05-31 | 河南工业大学 | A kind of enzyme process and the compound method for preparing dual network gel of heat treatment |
| CN106729934A (en) * | 2017-01-19 | 2017-05-31 | 上海交通大学 | A kind of IPN colloid of tomato skin soluble dietary fiber and preparation method thereof |
| CN107308505A (en) * | 2017-05-11 | 2017-11-03 | 华南理工大学 | A kind of sodium alginate-modified gelatin modified dopamine compound rest and preparation method thereof |
| CN107312193A (en) * | 2017-06-27 | 2017-11-03 | 吉林大学 | A kind of bionical injectable adhesion hydrogel, preparation method and its application in terms of biology |
| CN107916260A (en) * | 2017-09-29 | 2018-04-17 | 浙江大学 | One kind wraps up single celled method and product and application based on hydrogel |
| CN108126243A (en) * | 2017-04-04 | 2018-06-08 | 广州普钰智科技有限公司 | A kind of quick-gelatinizing injectable multilayer gel stent and preparation method thereof |
| CN108310460A (en) * | 2018-02-02 | 2018-07-24 | 武汉大学 | Injectable high intensity Thermo-sensitive modified chitin based aquagel and its preparation method and application |
| CN108478852A (en) * | 2018-04-08 | 2018-09-04 | 湖北工程学院 | A kind of pharmaceutical products, the carrier for loading pharmaceutical products, hydrogel and its method |
| CN108587191A (en) * | 2018-05-18 | 2018-09-28 | 重庆科技学院 | A kind of silk-fibroin/hyaluronic acid interpenetration network hydrogel and preparation method thereof |
| CN108778258A (en) * | 2015-12-18 | 2018-11-09 | 食品研究发展中心有限公司 | The covalent matrix of biodegradable of oral insulin for providing targeting colon activated by micropopulation and the method for obtaining these matrix |
| CN108882951A (en) * | 2016-02-12 | 2018-11-23 | 罗丹菲尔茨有限责任公司 | Moisture-keeping composition and application thereof |
| CN109053928A (en) * | 2018-05-30 | 2018-12-21 | 中国科学院兰州化学物理研究所 | A kind of large biological molecule and its preparation method and application based on modification of chitosan |
| CN109730976A (en) * | 2018-12-24 | 2019-05-10 | 华中科技大学 | A method of albumin nano granular is prepared based on free-radical oxidation |
| CN110152062A (en) * | 2019-05-22 | 2019-08-23 | 南通大学 | A kind of 3D biometric print hydrogel scaffold and preparation method thereof applied to regeneration |
| CN110464871A (en) * | 2019-08-21 | 2019-11-19 | 南开大学 | A kind of injection aquagel and preparation method thereof |
| CN110483804A (en) * | 2019-05-24 | 2019-11-22 | 深圳先进技术研究院 | Modified biopolymer and its application in 3D printing |
| CN110527116A (en) * | 2019-09-17 | 2019-12-03 | 江南大学 | A kind of method that enzyme process prepares the compound outer compress material of fibroin/hyaluronic acid |
| CN110540661A (en) * | 2019-09-30 | 2019-12-06 | 湖北工程学院 | Composite hydrogel of silk fibroin and polyvinyl alcohol, and preparation method and application thereof |
| CN110628090A (en) * | 2019-10-17 | 2019-12-31 | 陕西科技大学 | Cationic guar gum/chitosan composite hydrogel and preparation method thereof |
| CN110760103A (en) * | 2019-11-12 | 2020-02-07 | 四川大学 | Viscoelastic hydrogel and preparation method and application thereof |
| CN110772477A (en) * | 2019-11-05 | 2020-02-11 | 深圳市天达康基因工程有限公司 | Recombinant adenovirus sustained-release hydrogel, preparation method and application thereof |
| CN110982021A (en) * | 2018-10-03 | 2020-04-10 | 明基材料股份有限公司 | Colorant compositions for tinting silicone hydrogel contact lenses, contact lenses containing same, and methods of modifying silicone hydrogel contact lens colorants |
| CN111440340A (en) * | 2020-04-30 | 2020-07-24 | 苏州苏豪生物材料科技有限公司 | Silk fibroin-sodium hyaluronate cross-linked double-network gel and preparation method thereof |
| CN111568855A (en) * | 2020-06-08 | 2020-08-25 | 山东大学 | Preparation method of injectable hydrogel and application of injectable hydrogel in postoperative tumor treatment |
| CN111603617A (en) * | 2015-04-03 | 2020-09-01 | 伊诺特纳皮株式会社 | Hemostatic injection needle coated with cross-linked chitosan having catechol groups and oxidized catechol groups |
| CN111705512A (en) * | 2020-07-01 | 2020-09-25 | 徐州光彩丝绵制品有限公司 | Silk floss finishing method |
| CN111701074A (en) * | 2020-06-01 | 2020-09-25 | 温州医科大学 | Polydopamine hybrid agarose hydrogel, and preparation method and application thereof |
| CN112159532A (en) * | 2020-07-03 | 2021-01-01 | 中国科学院大学温州研究院(温州生物材料与工程研究所) | Oxygen-containing hydrogel dressing and preparation and application thereof |
| CN112494710A (en) * | 2019-09-16 | 2021-03-16 | 天津大学 | Transglutamineenzymatic crosslinked double-network adhesive and preparation method thereof |
| CN112500528A (en) * | 2020-12-03 | 2021-03-16 | 山东省眼科研究所 | Drug-loaded contact lens with antibacterial effect and preparation method thereof |
| CN113209365A (en) * | 2021-05-31 | 2021-08-06 | 福州大学 | Multifunctional closed hemostatic wound dressing and preparation method thereof |
| CN113244437A (en) * | 2021-06-15 | 2021-08-13 | 福州大学 | Preparation method and application of bionic multifunctional composite sponge dressing |
| CN113563681A (en) * | 2021-07-16 | 2021-10-29 | 南京大学 | Degradable wet-state adhesive hydrogel material and preparation method and application thereof |
| CN113769156A (en) * | 2021-08-26 | 2021-12-10 | 四川大学 | Hybrid fiber sponge with functions of hemostasis and wound repair and preparation method thereof |
| CN113924132A (en) * | 2019-05-07 | 2022-01-11 | 罗盖特公司 | Novel polysaccharide-based hydrogel scaffold for wound care |
| CN113956413A (en) * | 2021-11-11 | 2022-01-21 | 南京工业大学 | Preparation method of nano composite hydrogel and application of nano composite hydrogel in promotion of diabetic wound healing |
| CN114392728A (en) * | 2021-12-29 | 2022-04-26 | 重庆大学 | Magnetic hydrogel capable of efficiently adsorbing cationic dye and preparation method and application thereof |
| CN114404651A (en) * | 2022-01-25 | 2022-04-29 | 中科南京绿色制造产业创新研究院 | Calcium carbonate compounded dipeptide hydrogel scaffold and preparation method and application thereof |
| CN114835917A (en) * | 2022-04-13 | 2022-08-02 | 西南大学 | Preparation method of regenerated silk fibroin/epsilon-polylysine composite hydrogel, product and application thereof |
| CN114908052A (en) * | 2022-04-27 | 2022-08-16 | 华南理工大学 | Kit for culturing tumor stem cells and application thereof |
| CN115105628A (en) * | 2022-07-22 | 2022-09-27 | 广州市暨生元生物科技有限公司 | High-stability recombinant human epidermal growth factor wound repair dressing and preparation method thereof |
| CN115887772A (en) * | 2022-11-17 | 2023-04-04 | 南京工业大学 | Gelatin/sodium alginate hydrogel-based 3D printing biological ink and application thereof |
| CN115957381A (en) * | 2023-01-30 | 2023-04-14 | 复旦大学附属中山医院 | Injectable sericin hydrogel for regulating bone immune microenvironment |
| CN116870241A (en) * | 2023-08-03 | 2023-10-13 | 海南大学 | In-situ formed double-network hydrogel dressing and preparation method and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999016893A2 (en) * | 1997-09-29 | 1999-04-08 | Instituut Voor Agrotechnologisch Onderzoek (Ato-Dlo) | Enzymatic modification |
| CN101052684A (en) * | 2004-07-09 | 2007-10-10 | 克利夫兰临床基金会 | Hydroxyphenyl cross-linked macromolecular network and applications thereof |
| CN101439206A (en) * | 2007-11-22 | 2009-05-27 | 郭倩 | Preparation of enzyme-catalyzed rapid-solidified hydrogel and use thereof |
| WO2009148405A1 (en) * | 2008-06-05 | 2009-12-10 | Agency For Science, Technology And Research | Formation of hydrogel in the presence of peroxidase and low concentration of hydrogen peroxide |
| CN102532566A (en) * | 2011-12-21 | 2012-07-04 | 四川大学 | Preparation method of interpenetrating network composite hydrogel |
| CN102688525A (en) * | 2012-05-07 | 2012-09-26 | 东南大学 | Bio-macromolecular hydrogel and preparation method thereof |
-
2014
- 2014-06-12 CN CN201410259494.5A patent/CN104004231A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1999016893A2 (en) * | 1997-09-29 | 1999-04-08 | Instituut Voor Agrotechnologisch Onderzoek (Ato-Dlo) | Enzymatic modification |
| CN101052684A (en) * | 2004-07-09 | 2007-10-10 | 克利夫兰临床基金会 | Hydroxyphenyl cross-linked macromolecular network and applications thereof |
| CN101439206A (en) * | 2007-11-22 | 2009-05-27 | 郭倩 | Preparation of enzyme-catalyzed rapid-solidified hydrogel and use thereof |
| WO2009148405A1 (en) * | 2008-06-05 | 2009-12-10 | Agency For Science, Technology And Research | Formation of hydrogel in the presence of peroxidase and low concentration of hydrogen peroxide |
| CN102532566A (en) * | 2011-12-21 | 2012-07-04 | 四川大学 | Preparation method of interpenetrating network composite hydrogel |
| CN102688525A (en) * | 2012-05-07 | 2012-09-26 | 东南大学 | Bio-macromolecular hydrogel and preparation method thereof |
Non-Patent Citations (2)
| Title |
|---|
| 胡喜斌: "《动物药理》", 31 January 2013, 中国轻工业出版社 * |
| 薛巍等: "《生物医用水凝胶》", 31 December 2012, 暨南大学出版社 * |
Cited By (76)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104667349B (en) * | 2015-02-06 | 2017-01-25 | 福州大学 | Growth factor-loading silk fibroin/collagen bracket material and preparation method thereof |
| CN104667349A (en) * | 2015-02-06 | 2015-06-03 | 福州大学 | Growth factor-loading silk fibroin/collagen bracket material and preparation method thereof |
| CN111603617B (en) * | 2015-04-03 | 2022-05-24 | 伊诺特纳皮株式会社 | Hemostatic injection needle coated with cross-linked chitosan having catechol groups and oxidized catechol groups |
| CN111603617A (en) * | 2015-04-03 | 2020-09-01 | 伊诺特纳皮株式会社 | Hemostatic injection needle coated with cross-linked chitosan having catechol groups and oxidized catechol groups |
| CN105131632A (en) * | 2015-09-21 | 2015-12-09 | 高瑞杰 | Environment-friendly food package film based on modified elastic aloe gel micro powder and preparation method |
| CN105254906B (en) * | 2015-11-13 | 2017-11-03 | 东南大学 | Chemical crosslinking and the preparation method of the interpenetration network hydrogel of complexing of metal ion |
| CN105254906A (en) * | 2015-11-13 | 2016-01-20 | 东南大学 | Method for preparing chemical crosslinking and metal ion complexing type interpenetrating network hydrogel |
| CN108778258A (en) * | 2015-12-18 | 2018-11-09 | 食品研究发展中心有限公司 | The covalent matrix of biodegradable of oral insulin for providing targeting colon activated by micropopulation and the method for obtaining these matrix |
| CN108778258B (en) * | 2015-12-18 | 2023-02-28 | 食品研究发展中心有限公司 | Microbiota activated biodegradable covalent matrices for providing oral insulin targeting the colon and methods of obtaining these matrices |
| CN108882951B (en) * | 2016-02-12 | 2022-01-11 | 罗丹菲尔茨有限责任公司 | Moisturizing composition and application thereof |
| CN108882951A (en) * | 2016-02-12 | 2018-11-23 | 罗丹菲尔茨有限责任公司 | Moisture-keeping composition and application thereof |
| CN105944150B (en) * | 2016-03-25 | 2019-05-10 | 华东理工大学 | A kind of hydrogel and the artificial skin and preparation method thereof using the hydrogel |
| CN105944150A (en) * | 2016-03-25 | 2016-09-21 | 华东理工大学 | Hydrogel, artificial skin manufactured through application of hydrogel and manufacturing method of artificial skin |
| CN105802251A (en) * | 2016-04-25 | 2016-07-27 | 成都大学 | Self-assembly collagen template tissue engineering material as well as preparation method and application thereof |
| CN105802251B (en) * | 2016-04-25 | 2019-09-17 | 成都大学 | A kind of self assembly collagen template tissue engineering material and the preparation method and application thereof |
| CN106179272A (en) * | 2016-08-10 | 2016-12-07 | 合众（佛山）化工有限公司 | A kind of novel porous nano carbon microsphere composite aquogel and preparation method thereof |
| CN106750393A (en) * | 2016-11-15 | 2017-05-31 | 河南工业大学 | A kind of enzyme process and the compound method for preparing dual network gel of heat treatment |
| CN106700099A (en) * | 2016-11-23 | 2017-05-24 | 华中科技大学同济医学院附属协和医院 | Colorless and transparent silk sericin hydrogel, stent prepared therefrom and application thereof |
| CN106729934A (en) * | 2017-01-19 | 2017-05-31 | 上海交通大学 | A kind of IPN colloid of tomato skin soluble dietary fiber and preparation method thereof |
| CN106729934B (en) * | 2017-01-19 | 2020-03-20 | 上海交通大学 | Interpenetrating colloid of tomato peel soluble dietary fiber and preparation method thereof |
| CN108126243A (en) * | 2017-04-04 | 2018-06-08 | 广州普钰智科技有限公司 | A kind of quick-gelatinizing injectable multilayer gel stent and preparation method thereof |
| CN107308505A (en) * | 2017-05-11 | 2017-11-03 | 华南理工大学 | A kind of sodium alginate-modified gelatin modified dopamine compound rest and preparation method thereof |
| CN107308505B (en) * | 2017-05-11 | 2021-02-12 | 华南理工大学 | Sodium alginate-modified gelatin-modified dopamine composite scaffold and preparation method thereof |
| CN107312193A (en) * | 2017-06-27 | 2017-11-03 | 吉林大学 | A kind of bionical injectable adhesion hydrogel, preparation method and its application in terms of biology |
| CN107916260A (en) * | 2017-09-29 | 2018-04-17 | 浙江大学 | One kind wraps up single celled method and product and application based on hydrogel |
| CN107916260B (en) * | 2017-09-29 | 2021-03-05 | 浙江大学 | Hydrogel-based single cell wrapping method, product and application |
| CN108310460B (en) * | 2018-02-02 | 2021-08-03 | 武汉大学 | Injectable high-strength temperature-sensitive modified chitin-based hydrogel and preparation method and application thereof |
| CN108310460A (en) * | 2018-02-02 | 2018-07-24 | 武汉大学 | Injectable high intensity Thermo-sensitive modified chitin based aquagel and its preparation method and application |
| CN108478852A (en) * | 2018-04-08 | 2018-09-04 | 湖北工程学院 | A kind of pharmaceutical products, the carrier for loading pharmaceutical products, hydrogel and its method |
| CN108587191A (en) * | 2018-05-18 | 2018-09-28 | 重庆科技学院 | A kind of silk-fibroin/hyaluronic acid interpenetration network hydrogel and preparation method thereof |
| CN109053928A (en) * | 2018-05-30 | 2018-12-21 | 中国科学院兰州化学物理研究所 | A kind of large biological molecule and its preparation method and application based on modification of chitosan |
| CN110982021A (en) * | 2018-10-03 | 2020-04-10 | 明基材料股份有限公司 | Colorant compositions for tinting silicone hydrogel contact lenses, contact lenses containing same, and methods of modifying silicone hydrogel contact lens colorants |
| CN110982021B (en) * | 2018-10-03 | 2022-03-08 | 明基材料股份有限公司 | Colorant compositions for tinting silicone hydrogel contact lenses, contact lenses containing same, and methods of modifying silicone hydrogel contact lens colorants |
| CN109730976A (en) * | 2018-12-24 | 2019-05-10 | 华中科技大学 | A method of albumin nano granular is prepared based on free-radical oxidation |
| CN109730976B (en) * | 2018-12-24 | 2020-06-09 | 华中科技大学 | Method for preparing albumin nanoparticles based on free radical oxidation |
| CN113924132B (en) * | 2019-05-07 | 2023-07-25 | 罗盖特公司 | Novel polysaccharide-based hydrogel scaffold for wound care |
| CN113924132A (en) * | 2019-05-07 | 2022-01-11 | 罗盖特公司 | Novel polysaccharide-based hydrogel scaffold for wound care |
| CN110152062A (en) * | 2019-05-22 | 2019-08-23 | 南通大学 | A kind of 3D biometric print hydrogel scaffold and preparation method thereof applied to regeneration |
| CN110483804A (en) * | 2019-05-24 | 2019-11-22 | 深圳先进技术研究院 | Modified biopolymer and its application in 3D printing |
| CN110464871A (en) * | 2019-08-21 | 2019-11-19 | 南开大学 | A kind of injection aquagel and preparation method thereof |
| CN112494710A (en) * | 2019-09-16 | 2021-03-16 | 天津大学 | Transglutamineenzymatic crosslinked double-network adhesive and preparation method thereof |
| CN112494710B (en) * | 2019-09-16 | 2021-12-14 | 天津大学 | Transglutamineenzymatic crosslinked double-network adhesive and preparation method thereof |
| CN110527116A (en) * | 2019-09-17 | 2019-12-03 | 江南大学 | A kind of method that enzyme process prepares the compound outer compress material of fibroin/hyaluronic acid |
| CN110540661B (en) * | 2019-09-30 | 2022-03-29 | 湖北工程学院 | Composite hydrogel of silk fibroin and polyvinyl alcohol, and preparation method and application thereof |
| CN110540661A (en) * | 2019-09-30 | 2019-12-06 | 湖北工程学院 | Composite hydrogel of silk fibroin and polyvinyl alcohol, and preparation method and application thereof |
| CN110628090B (en) * | 2019-10-17 | 2021-06-22 | 陕西科技大学 | Cationic guar gum/chitosan composite hydrogel and preparation method thereof |
| CN110628090A (en) * | 2019-10-17 | 2019-12-31 | 陕西科技大学 | Cationic guar gum/chitosan composite hydrogel and preparation method thereof |
| CN110772477A (en) * | 2019-11-05 | 2020-02-11 | 深圳市天达康基因工程有限公司 | Recombinant adenovirus sustained-release hydrogel, preparation method and application thereof |
| CN110760103A (en) * | 2019-11-12 | 2020-02-07 | 四川大学 | Viscoelastic hydrogel and preparation method and application thereof |
| CN110760103B (en) * | 2019-11-12 | 2021-09-14 | 四川大学 | Viscoelastic hydrogel and preparation method and application thereof |
| CN111440340B (en) * | 2020-04-30 | 2022-04-12 | 苏州苏豪生物材料科技有限公司 | Silk fibroin-sodium hyaluronate cross-linked double-network gel and preparation method thereof |
| CN111440340A (en) * | 2020-04-30 | 2020-07-24 | 苏州苏豪生物材料科技有限公司 | Silk fibroin-sodium hyaluronate cross-linked double-network gel and preparation method thereof |
| CN111701074A (en) * | 2020-06-01 | 2020-09-25 | 温州医科大学 | Polydopamine hybrid agarose hydrogel, and preparation method and application thereof |
| CN111568855A (en) * | 2020-06-08 | 2020-08-25 | 山东大学 | Preparation method of injectable hydrogel and application of injectable hydrogel in postoperative tumor treatment |
| CN111568855B (en) * | 2020-06-08 | 2022-02-08 | 山东大学 | Preparation method of injectable hydrogel and application of injectable hydrogel in postoperative tumor treatment |
| CN111705512A (en) * | 2020-07-01 | 2020-09-25 | 徐州光彩丝绵制品有限公司 | Silk floss finishing method |
| CN112159532A (en) * | 2020-07-03 | 2021-01-01 | 中国科学院大学温州研究院(温州生物材料与工程研究所) | Oxygen-containing hydrogel dressing and preparation and application thereof |
| CN112500528A (en) * | 2020-12-03 | 2021-03-16 | 山东省眼科研究所 | Drug-loaded contact lens with antibacterial effect and preparation method thereof |
| CN113209365A (en) * | 2021-05-31 | 2021-08-06 | 福州大学 | Multifunctional closed hemostatic wound dressing and preparation method thereof |
| CN113209365B (en) * | 2021-05-31 | 2022-04-01 | 福州大学 | Multifunctional closed hemostatic wound dressing and preparation method thereof |
| CN113244437A (en) * | 2021-06-15 | 2021-08-13 | 福州大学 | Preparation method and application of bionic multifunctional composite sponge dressing |
| CN113563681A (en) * | 2021-07-16 | 2021-10-29 | 南京大学 | Degradable wet-state adhesive hydrogel material and preparation method and application thereof |
| CN113769156A (en) * | 2021-08-26 | 2021-12-10 | 四川大学 | Hybrid fiber sponge with functions of hemostasis and wound repair and preparation method thereof |
| CN113956413A (en) * | 2021-11-11 | 2022-01-21 | 南京工业大学 | Preparation method of nano composite hydrogel and application of nano composite hydrogel in promotion of diabetic wound healing |
| CN114392728B (en) * | 2021-12-29 | 2023-02-03 | 重庆大学 | Magnetic hydrogel capable of efficiently adsorbing cationic dye and preparation method and application thereof |
| CN114392728A (en) * | 2021-12-29 | 2022-04-26 | 重庆大学 | Magnetic hydrogel capable of efficiently adsorbing cationic dye and preparation method and application thereof |
| CN114404651A (en) * | 2022-01-25 | 2022-04-29 | 中科南京绿色制造产业创新研究院 | Calcium carbonate compounded dipeptide hydrogel scaffold and preparation method and application thereof |
| CN114835917A (en) * | 2022-04-13 | 2022-08-02 | 西南大学 | Preparation method of regenerated silk fibroin/epsilon-polylysine composite hydrogel, product and application thereof |
| CN114835917B (en) * | 2022-04-13 | 2024-03-19 | 西南大学 | Preparation method of regenerated silk fibroin/epsilon-polylysine composite hydrogel, product and application thereof |
| CN114908052A (en) * | 2022-04-27 | 2022-08-16 | 华南理工大学 | Kit for culturing tumor stem cells and application thereof |
| CN114908052B (en) * | 2022-04-27 | 2023-12-19 | 华南理工大学 | Kit for culturing tumor stem cells and application thereof |
| CN115105628A (en) * | 2022-07-22 | 2022-09-27 | 广州市暨生元生物科技有限公司 | High-stability recombinant human epidermal growth factor wound repair dressing and preparation method thereof |
| CN115105628B (en) * | 2022-07-22 | 2024-03-19 | 广州市暨生元生物科技有限公司 | High-stability recombinant human epidermal growth factor wound repair dressing and preparation method thereof |
| CN115887772A (en) * | 2022-11-17 | 2023-04-04 | 南京工业大学 | Gelatin/sodium alginate hydrogel-based 3D printing biological ink and application thereof |
| CN115957381A (en) * | 2023-01-30 | 2023-04-14 | 复旦大学附属中山医院 | Injectable sericin hydrogel for regulating bone immune microenvironment |
| CN116870241A (en) * | 2023-08-03 | 2023-10-13 | 海南大学 | In-situ formed double-network hydrogel dressing and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN104004231A (en) | Biomacromolecule interpenetrating polymer network hydrogel and preparation method thereof | |
| CN102688525B (en) | Bio-macromolecular hydrogel and preparation method thereof | |
| Liu et al. | Thermosensitive injectable in-situ forming carboxymethyl chitin hydrogel for three-dimensional cell culture | |
| Agheb et al. | Novel electrospun nanofibers of modified gelatin-tyrosine in cartilage tissue engineering | |
| Guo et al. | Hydrogels of collagen/chondroitin sulfate/hyaluronan interpenetrating polymer network for cartilage tissue engineering | |
| Dahlmann et al. | Fully defined in situ cross-linkable alginate and hyaluronic acid hydrogels for myocardial tissue engineering | |
| Yan et al. | Injectable in situ self-cross-linking hydrogels based on poly (L-glutamic acid) and alginate for cartilage tissue engineering | |
| Hu et al. | Cell immobilization in gelatin–hydroxyphenylpropionic acid hydrogel fibers | |
| Jin et al. | Chondrogenesis in injectable enzymatically crosslinked heparin/dextran hydrogels | |
| Li et al. | Enzymatically crosslinked and mechanically tunable silk fibroin/pullulan hydrogels for mesenchymal stem cells delivery | |
| US9624259B2 (en) | Compositions and methods for scaffold formation | |
| US20170014543A1 (en) | HYDROGEL BASED ON γ-POLYGLUTAMIC ACID AND ε-POLYLYSINE CROSSLINKED POLYMER, AND PREPARATION METHOD THEREFOR | |
| KR20220077156A (en) | Biogum and botanical gum hydrogel bioinks for the physiological 3d bioprinting of tissue constructs for in vitro culture and transplantation | |
| CN1208094C (en) | Base materials for tissue regeneration, transplant materials and process for producing the same | |
| CN104262648A (en) | Collagen-based biomedical material by taking dialdehyde polyethylene glycol as cross-linking agent and preparation method thereof | |
| CN114524953A (en) | Silk fibroin/hyaluronic acid composite hydrogel, preparation method and application | |
| Wang et al. | Extracellular matrix mimicking dynamic interpenetrating network hydrogel for skin tissue engineering | |
| Yang et al. | Preparation and characterization of macromolecule cross-linked collagen hydrogels for chondrocyte delivery | |
| CN114874455A (en) | Construction method of neutral-dissolved modified collagen and gel with self-assembly capability and photo-crosslinking capability | |
| CN104784758B (en) | Preparation method of polymer/keratin composite anticoagulation vascular tissue engineering scaffold | |
| CN105176095B (en) | A kind of collagen-based organic silicon rubber porous composite film and preparation method thereof | |
| CN113980294B (en) | Sodium alginate-based conductive self-healing hydrogel and preparation method and application thereof | |
| Fan et al. | Biocompatible conjugation for biodegradable hydrogels as drug and cell scaffolds | |
| WO2004082728A1 (en) | Medical material | |
| FR3035107A1 (en) | PROCESS FOR PREPARING HYDROGEL FROM MODIFIED HYALURONIC ACID AND TYPE I COLLAGEN |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140827 |