CN110229530B - Double-network silk fibroin hydrogel and preparation method thereof - Google Patents
Double-network silk fibroin hydrogel and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a double-network silk fibroin hydrogel and a preparation method thereof. The invention takes silk fibroin macromolecules with good biocompatibility as a matrix, introduces a host and an object through chemical modification, and introduces horse radish peroxidase and hydrogen peroxide into a system to prepare the double-network silk fibroin hydrogel. Reversible binding between the host and the guest unit and cross-linking catalyzed by horseradish peroxidase endow the silk fibroin hydrogel with structural and performance stability. Meanwhile, the existence of the double-network structure increases the crosslinking action sites of the gel network, greatly improves the elastic modulus of the silk fibroin hydrogel and further improves the tensile property of the silk fibroin hydrogel.
Description
Technical Field
The invention relates to the technical field of biological materials, in particular to a double-network silk fibroin hydrogel and a preparation method thereof.
Background
The hydrogel is one of soft materials, is formed by crosslinking hydrophilic polymer chains in water, has high water content, can transport oxygen and nutrients, is similar to natural extracellular matrix, and has wide application prospects in the fields of tissue engineering, drug delivery, regenerative medicine and the like. Silk fibroin is natural polymer fiber protein extracted from silk, and is approved by FDA and can be used for clinical medical treatment. The silk fibroin-based hydrogel has excellent biocompatibility and biodegradability, can provide a proper microenvironment for proliferation and differentiation of cells, can realize repair and regeneration of tissues, and has important application value in the field of biomedicine. The silk fibroin-based hydrogel can be prepared by methods of ultrasound, vortex, electric field, pH reduction and the like, but the prepared gel has weaker mechanical property, does not have the elasticity and toughness of biological tissues, has poor plastic deformation when the strain is higher than 10 percent, and is easy to damage and break along with the time extension, thereby greatly limiting the application of the silk fibroin-based hydrogel in the field of biological medicines. The plastic deformation of the silk fibroin hydrogel reported in the prior art is poor when the strain is higher than 10%, the material is easy to be damaged and broken, and the repeated tensile property is poor, and how to improve the toughness strength of the silk fibroin hydrogel is an important challenge in hydrogel research.
Disclosure of Invention
The invention aims to provide a double-network silk fibroin hydrogel and a preparation method thereof, so as to overcome the defect of poor tensile property of the traditional silk fibroin hydrogel.
Therefore, the invention provides a double-network silk fibroin hydrogel which is constructed by taking silk fibroin molecules as a matrix and introducing a host, an object and horseradish peroxidase into an amino acid side chain of the matrix.
Preferably, the host comprises cyclodextrin or cucurbituril, and the guest comprises adamantane, ferrocene, cholic acid, cholesterol or carbazole derivatives.
The invention also provides a preparation method of the double-network silk fibroin hydrogel, which comprises the following steps:
(1) preparing carboxylated silk fibroin powder;
(2) preparing a silk fibroin-host solution;
(3) preparing a silk fibroin-guest solution;
(4) adding a horseradish peroxidase solution into the silk fibroin-main body solution, and uniformly mixing to obtain a first mixed solution;
(5) adding a hydrogen peroxide solution into the silk fibroin-object solution, and uniformly mixing to obtain a second mixed solution;
(6) and uniformly mixing the first mixed solution and the second mixed solution, and standing to obtain the double-network silk fibroin hydrogel.
Preferably, the step (1) includes: degumming, dissolving and dialyzing silk to obtain a silk fibroin solution; carrying out chemical modification on the silk fibroin solution by borax, diazonium salt and chloroacetic acid to obtain 0.1-50% carboxylated silk fibroin solution; and dialyzing the carboxylated silk fibroin solution, and freeze-drying to obtain carboxylated silk fibroin powder.
Preferably, the step (2) includes: dissolving the aminated main body in dimethyl sulfoxide solution to prepare aminated main body solution with concentration of 0.1-1000 mg/mL; dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with concentration of 0.1-1000 mg/mL; dissolving carboxylated silk fibroin powder in dimethyl sulfoxide solution to prepare carboxylated silk fibroin solution with the concentration of 0.05-10 mg/mL; mixing the aminated main body solution, an EDC.HCl solution and a carboxylated silk fibroin solution, reacting at room temperature, dialyzing, freezing and drying to obtain silk fibroin-main body powder; dissolving the silk fibroin-main body powder in water to prepare a silk fibroin-main body solution with the mass fraction of 1-50%.
Preferably, the step (3) includes: preparing an object solution with the concentration of 0.1-1000 mg/mL; dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with concentration of 0.1-1000 mg/mL; dissolving carboxylated silk fibroin powder in dimethyl sulfoxide solution to prepare carboxylated silk fibroin solution with the concentration of 0.05-10 mg/mL; mixing the object solution, an EDC.HCl solution and a carboxylated silk fibroin solution, reacting at room temperature, dialyzing, freezing and drying to obtain silk fibroin-object powder; dissolving silk fibroin-object powder in water to prepare silk fibroin-object solution with the mass fraction of 1% -50%.
Preferably, in the step (4), the content of the horseradish peroxidase is 900-1100U/mL.
Preferably, in the step (5), the concentration of the hydrogen peroxide solution is 160-170 mM.
Preferably, in the step (6), the first mixed solution and the second mixed solution are uniformly mixed according to the molar ratio of the host to the guest of 100:1-1:100, and the mass fraction of the double-network silk fibroin hydrogel is 1% -50%.
Compared with the prior art, the invention has the advantages and positive effects that: the invention provides a double-network silk fibroin hydrogel and a preparation method thereof. The invention takes silk fibroin macromolecules with good biocompatibility as a matrix, introduces a host and an object through chemical modification, and introduces horse radish peroxidase and hydrogen peroxide into a system to prepare the double-network silk fibroin hydrogel. Reversible binding between the host and the guest unit and cross-linking catalyzed by horseradish peroxidase endow the silk fibroin hydrogel with structural and performance stability. Meanwhile, the existence of the double-network structure increases the crosslinking action sites of the gel network, greatly improves the elastic modulus of the silk fibroin hydrogel and further improves the tensile property of the silk fibroin hydrogel. The silk fibroin hydrogel prepared by the invention has the advantages of softness, high water content, bioactivity, strong toughness and tensile resistance, is not easy to damage and break, can withstand tensile strain deformation of 800 percent, realizes repeated high-strength stretching within the strain range of 800 percent, and can quickly recover the original mechanical property after tensile breaking. The double-network silk fibroin hydrogel provides a new idea for designing and preparing composite materials for biological tissue engineering, and expands the practical application of the silk fibroin hydrogel in the fields of biomedicine, tissue regeneration, flexible electronic devices and the like.
Other features and advantages of the present invention will become more apparent upon reading of the following detailed description of the invention in conjunction with the accompanying drawings.
Drawings
FIG. 1 is an optical photograph of a double-network silk fibroin hydrogel of example 3 of the present invention before and after stretching;
fig. 2 is a tensile stress-strain curve of the double-network silk fibroin hydrogel of example 4 of the present invention.
Detailed Description
The following detailed description of specific embodiments of the present invention is provided to illustrate and explain the present invention and to be understood not to limit the present invention.
The invention provides a double-network silk fibroin hydrogel which is constructed by taking silk fibroin molecules as a matrix and introducing a host, an object and horseradish peroxidase into an amino acid side chain of the silk fibroin molecules.
The host can be cyclodextrin or cucurbituril, and the cyclodextrin and the cucurbituril both have a macrocyclic molecular structure and can form an inclusion complex with a guest molecule. The guest can be adamantane, ferrocene, cholic acid, cholesterol or carbazole derivatives, and can form an inclusion complex with the host molecule.
The host is preferably cyclodextrin and the guest is preferably adamantane.
The invention takes silk fibroin macromolecules with good biocompatibility as a matrix, introduces a host and an object through chemical modification, and introduces horse radish peroxidase and hydrogen peroxide into a system to prepare the double-network silk fibroin hydrogel. Reversible binding between the host and the guest unit and cross-linking catalyzed by horseradish peroxidase endow the silk fibroin hydrogel with structural and performance stability. Meanwhile, the existence of the double-network structure increases the crosslinking action sites of the gel network, greatly improves the elastic modulus of the silk fibroin hydrogel and further improves the tensile property of the silk fibroin hydrogel. The silk fibroin hydrogel prepared by the invention has the advantages of softness, high water content, bioactivity, strong toughness and tensile resistance, is not easy to damage and break, can withstand tensile strain deformation of 800 percent, realizes repeated high-strength stretching within the strain range of 800 percent, and can quickly recover the original mechanical property after tensile breaking. The double-network silk fibroin hydrogel provides a new idea for designing and preparing composite materials for biological tissue engineering, and expands the practical application of the silk fibroin hydrogel in the fields of biomedicine, tissue regeneration, flexible electronic devices and the like.
According to the invention, the stretching performance of the silk fibroin gel is improved by combining two crosslinking modes of physical crosslinking and chemical crosslinking, firstly, a host and an object are grafted on a silk fibroin macromolecular chain in a chemical modification mode, and the host and the object form an inclusion complex to form a physical crosslinking network, so that the silk fibroin gel with excellent stretching performance is obtained. Secondly, hydrogen peroxide can react with Fe (III) in horseradish peroxidase to generate an intermediate (HRP I) with strong oxidizing property, phenolic hydroxyl on the silk fibroin tyrosine forms phenolic free radicals under the oxidizing action of the HRP I, and then the phenolic free radicals on the tyrosine form a covalent bond of dicarbonic acid under the catalytic action of the horseradish peroxidase to form a chemical crosslinking network, so that the tensile property of the silk fibroin hydrogel is further improved. The physical crosslinking effect between the subject and the object and the chemical crosslinking effect between horseradish peroxidase and hydrogen peroxide improve the elastic modulus of the silk fibroin gel, so that the silk fibroin gel has excellent tensile deformation capability.
The preparation method of the double-network silk fibroin hydrogel comprises the following steps:
(1) preparing a carboxylated silk fibroin solution; degumming, dissolving and dialyzing silk to obtain a silk fibroin solution; carrying out chemical modification on the silk fibroin solution by borax, diazonium salt and chloroacetic acid to obtain a carboxylated silk fibroin solution with the concentration of 0.1-50%; and dialyzing the carboxylated silk fibroin solution, and freeze-drying to obtain carboxylated silk fibroin powder. The concentration of the carboxylated silk fibroin solution is 0.1% -50%, and within the concentration range, the silk fibroin can be grafted with the cyclodextrin and the adamantane serving as the object to the greatest extent.
(2) Preparing a silk fibroin-host solution; dissolving the aminated main body in dimethyl sulfoxide solution to prepare aminated main body solution with concentration of 0.1-1000 mg/mL; dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with concentration of 0.1-1000 mg/mL; dissolving carboxylated silk fibroin powder in dimethyl sulfoxide solution to prepare carboxylated silk fibroin solution with the concentration of 0.05-10 mg/mL; mixing the aminated main body solution, an EDC.HCl solution and a carboxylated silk fibroin solution, reacting at room temperature, dialyzing, freezing and drying to obtain silk fibroin-main body powder; dissolving silk fibroin-main body powder in water to prepare a silk fibroin-main body solution with the mass fraction of 1-50%; within this concentration range, the host cyclodextrin can be maximally grafted with silk fibroin.
(3) Preparing a silk fibroin-guest solution; preparing an object solution with the concentration of 0.1-1000 mg/mL; dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with concentration of 0.1-1000 mg/mL; dissolving carboxylated silk fibroin powder in dimethyl sulfoxide solution to prepare carboxylated silk fibroin solution with the concentration of 0.05-10 mg/mL; mixing the object solution, an EDC.HCl solution and a carboxylated silk fibroin solution, reacting at room temperature, dialyzing, freezing and drying to obtain silk fibroin-object powder; dissolving silk fibroin-object powder in water to prepare a silk fibroin-object solution with the mass fraction of 1% -50%; within this concentration range, guest adamantane can be maximally grafted with silk fibroin.
(4) Adding 900-.
(5) Adding 160-170mM hydrogen peroxide solution into the silk fibroin-object solution, and uniformly mixing to obtain a second mixed solution. The concentration of the hydrogen peroxide solution is 160-170mM, and the hydrogen peroxide in the concentration range has the best catalytic effect on the enzyme.
(6) And uniformly mixing the first mixed solution and the second mixed solution according to the molar ratio of cyclodextrin to adamantane of 100:1-1:100, and standing to obtain the double-network silk fibroin hydrogel with the mass fraction of 1% -50%. When the first mixed solution and the second mixed solution are mixed according to the molar ratio of the cyclodextrin to the adamantane of 100:1-1:100, the performance of the formed double-network silk fibroin hydrogel is best, and the stretchability and the adhesion performance of the silk fibroin hydrogel can be improved to the greatest extent.
Example 1
(1) Preparing a carboxylated silk fibroin solution;
processing silk with Na2CO3Degumming the solution, dissolving the LiBr solution, and dialyzing with deionized water to obtain a silk fibroin solution with the concentration of 1%; mixing 5 mL silk fibroin solution with 2 mL, 10M NaOH solution, stirring for 5 min, adding 4 mL, 1M chloroacetic acid into the reaction solution, mixing at room temperature for 50 min, and adding 500 μ L, 7 mg/mL NaH into the above mixed solution2PO4The solution is prepared by mixing a solvent and a solvent,carrying out ice-bath reaction for 30 min to obtain a carboxylated silk fibroin solution; dialyzing, and freeze-drying to obtain carboxylated silk fibroin powder.
(2) Preparing a silk fibroin-cyclodextrin solution;
dissolving aminated cyclodextrin in dimethyl sulfoxide solution to prepare aminated cyclodextrin solution with concentration of 1 mg/mL; dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with the concentration of 1 mg/mL; dissolving 1 mg of carboxylated silk fibroin powder in 10mL of dimethyl sulfoxide solution, adding 1 mu L of aminated cyclodextrin solution and 1 mu L of EDC.HCl solution, reacting at room temperature for 24 h, dialyzing, freeze-drying, and preparing silk fibroin-cyclodextrin powder; dissolving silk fibroin-cyclodextrin powder in water to prepare a silk fibroin-cyclodextrin solution with the mass fraction of 1%.
(3) Preparing a silk fibroin-adamantane solution;
dissolving amantadine hydrochloride in a dimethyl sulfoxide solution to prepare an amantadine hydrochloride solution with the concentration of 1 mg/mL; dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with the concentration of 1 mg/mL; dissolving 1 mg of carboxylated silk fibroin in 10mL of dimethyl sulfoxide solution, adding 1 mu L of amantadine hydrochloride solution and 1 mu L of EDC.HCl, reacting at room temperature for 24 h, dialyzing, freeze-drying, and preparing silk fibroin-adamantane powder; dissolving silk fibroin-adamantane powder in water to prepare a silk fibroin-adamantane solution with the mass fraction of 1%.
(4) Adding 1000U/mL horseradish peroxidase solution into 10mL 1% silk fibroin-cyclodextrin solution, and uniformly mixing to obtain a first mixed solution.
(5) To 100. mu.L of 1% silk fibroin-adamantane solution, 165 mM hydrogen peroxide was added and mixed uniformly to obtain a second mixed solution.
(6) And uniformly mixing the first mixed solution and the second mixed solution according to the molar ratio of cyclodextrin to adamantane of 100:1, and standing to obtain the double-network silk fibroin hydrogel with the mass fraction of 1%.
Example 2
(1) Preparing a carboxylated silk fibroin solution;
processing silk with Na2CO3Degumming the solution, dissolving the LiBr solution, and dialyzing with deionized water to obtain a silk fibroin solution with the concentration of 4%; mixing 5 mL silk fibroin solution with 2 mL, 10M NaOH solution, stirring for 5 min, adding 4 mL, 1M chloroacetic acid into the reaction solution, mixing at room temperature for 50 min, and adding 500 μ L, 7 mg/mL NaH into the above mixed solution2PO4Carrying out ice-bath reaction on the solution for 30 min to obtain a carboxylated silk fibroin solution; dialyzing, and freeze-drying to obtain carboxylated silk fibroin powder.
(2) Preparing a silk fibroin-cyclodextrin solution;
dissolving aminated cyclodextrin in dimethyl sulfoxide solution to prepare aminated cyclodextrin solution with concentration of 10 mg/mL; respectively dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with the concentration of 10 mg/mL; dissolving 3 mg of carboxylated silk fibroin powder in 30 mL of dimethyl sulfoxide solution, adding 100 mu L of aminated cyclodextrin solution and 100 mu L of EDC.HCl solution, reacting at room temperature for 24 h, dialyzing, freeze-drying, and preparing silk fibroin-cyclodextrin powder; dissolving silk fibroin-cyclodextrin powder in water to prepare a silk fibroin-cyclodextrin solution with the mass fraction of 15%.
(3) Preparing a silk fibroin-adamantane solution;
dissolving amantadine hydrochloride in a dimethyl sulfoxide solution to prepare an amantadine hydrochloride solution with the concentration of 10 mg/mL; dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with the concentration of 10 mg/mL; dissolving 30 mg of carboxylated silk fibroin in 300 mL of dimethyl sulfoxide solution, adding 200 mu L of amantadine hydrochloride solution and 200 mu L of EDC.HCl, reacting at room temperature for 24 h, dialyzing, freeze-drying, and preparing silk fibroin-adamantane powder; dissolving silk fibroin-adamantane powder in water to prepare a silk fibroin-adamantane solution with the mass fraction of 1%.
(4) Adding 1000U/mL horseradish peroxidase solution into 10mL 15% silk fibroin-cyclodextrin solution, and uniformly mixing to obtain a first mixed solution.
(5) To 3 mL, 1% silk fibroin-adamantane solution was added 165 mM hydrogen peroxide, and mixed well to obtain a second mixed solution.
(6) And uniformly mixing the first mixed solution and the second mixed solution according to the molar ratio of cyclodextrin to adamantane of 50:1, and standing to obtain the double-network silk fibroin hydrogel with the mass fraction of 11.8%.
Example 3
(1) Preparing a carboxylated silk fibroin solution;
processing silk with Na2CO3Degumming the solution, dissolving the LiBr solution, and dialyzing with deionized water to obtain a 7% silk fibroin solution; mixing 5 mL silk fibroin solution with 2 mL, 10M NaOH solution, stirring for 5 min, adding 4 mL, 1M chloroacetic acid into the reaction solution, mixing at room temperature for 50 min, and adding 1 mL, 7 mg/mL NaH into the above mixed solution2PO4Carrying out ice-bath reaction on the solution for 30 min to obtain a carboxylated silk fibroin solution; dialyzing, and freeze-drying to obtain carboxylated silk fibroin powder.
(2) Preparing a silk fibroin-cyclodextrin solution;
dissolving aminated cyclodextrin in dimethyl sulfoxide solution to prepare aminated cyclodextrin solution with concentration of 100 mg/mL; respectively dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with the concentration of 100 mg/mL; dissolving 30 mg of carboxylated silk fibroin powder in 300 mL of dimethyl sulfoxide solution, adding 1000 mu L of aminated cyclodextrin solution and 100 mu L of EDC.HCl solution, reacting at room temperature for 24 h, dialyzing, freeze-drying, and preparing silk fibroin-cyclodextrin powder; dissolving silk fibroin-cyclodextrin powder in water to prepare a silk fibroin-cyclodextrin solution with the mass fraction of 25%.
(3) Preparing a silk fibroin-adamantane solution;
dissolving amantadine hydrochloride in a dimethyl sulfoxide solution to prepare an amantadine hydrochloride solution with the concentration of 100 mg/mL; dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with the concentration of 100 mg/mL; dissolving 30 mg of carboxylated silk fibroin in 300 mL of dimethyl sulfoxide solution, adding 200 mu L of amantadine hydrochloride solution and 200 mu L of EDC.HCl, reacting at room temperature for 24 h, dialyzing, freeze-drying, and preparing silk fibroin-adamantane powder; dissolving silk fibroin-adamantane powder in water to prepare a silk fibroin-adamantane solution with the mass fraction of 25%.
(4) Adding 1000U/mL horseradish peroxidase solution into 10mL, 25% silk fibroin-cyclodextrin solution, and mixing uniformly to obtain a first mixed solution.
(5) To 10mL, 25% silk fibroin-adamantane solution was added 165 mM hydrogen peroxide, and mixed well to obtain a second mixed solution.
(6) And uniformly mixing the first mixed solution and the second mixed solution according to the molar ratio of cyclodextrin to adamantane of 1:1, and standing to obtain the double-network silk fibroin hydrogel with the mass fraction of 25%.
Fig. 1 is an optical photograph of the double-network silk fibroin hydrogel of example 3 before and after stretching, which shows that the double-network silk fibroin hydrogel of this example has good stretchability, and can be stretched to 2.5 times of the original length.
Example 4
(1) Preparing a carboxylated silk fibroin solution;
processing silk with Na2CO3Degumming the solution, dissolving the LiBr solution, and dialyzing with deionized water to obtain a silk fibroin solution with the concentration of 10%; mixing 5 mL silk fibroin solution with 4 mL, 10M NaOH solution, stirring for 5 min, adding 4 mL, 2M chloroacetic acid into the reaction solution, mixing at room temperature for 50 min, and adding 1 mL, 7 mg/mL NaH into the above mixed solution2PO4Carrying out ice-bath reaction on the solution for 30 min to obtain a carboxylated silk fibroin solution; dialyzing, and freeze-drying to obtain carboxylated silk fibroin powder.
(2) Preparing a silk fibroin-cyclodextrin solution;
dissolving aminated cyclodextrin in dimethyl sulfoxide solution to prepare aminated cyclodextrin solution with concentration of 1000 mg/mL; respectively dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with the concentration of 1000 mg/mL; dissolving 30 mg of carboxylated silk fibroin powder in 300 mL of dimethyl sulfoxide solution, adding 1000 mu L of aminated cyclodextrin solution and 100 mu L of EDC.HCl solution, reacting at room temperature for 24 h, dialyzing, freeze-drying, and preparing silk fibroin-cyclodextrin powder; dissolving silk fibroin-cyclodextrin powder in water to prepare a silk fibroin-cyclodextrin solution with the mass fraction of 4%.
(3) Preparing a silk fibroin-adamantane solution;
dissolving amantadine hydrochloride in a dimethyl sulfoxide solution to prepare an amantadine hydrochloride solution with the concentration of 1000 mg/mL; dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with the concentration of 50 mg/mL; dissolving 30 mg of carboxylated silk fibroin in 300 mL of dimethyl sulfoxide solution, adding 200 mu L of amantadine hydrochloride solution and 200 mu L of EDC.HCl, reacting at room temperature for 24 h, dialyzing, freeze-drying, and preparing silk fibroin-adamantane powder; dissolving silk fibroin-adamantane powder in water to prepare a silk fibroin-adamantane solution with the mass fraction of 40%.
(4) Adding 1000U/mL horseradish peroxidase solution into 2 mL, 4% silk fibroin-cyclodextrin solution, and mixing uniformly to obtain a first mixed solution.
(5) To 20 mL, 40% silk fibroin-adamantane solution was added 165 mM hydrogen peroxide, and mixed well to obtain a second mixed solution.
(6) And uniformly mixing the first mixed solution and the second mixed solution according to the molar ratio of cyclodextrin to adamantane of 1:100, and standing to obtain the double-network silk fibroin hydrogel with the mass fraction of 36.7%.
Fig. 2 is a tensile stress-strain curve of the double-network silk fibroin hydrogel of example 4, which illustrates that the double-network silk fibroin hydrogel prepared by this example has very excellent tensile properties, and the tensile elongation is as high as 800%.
The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions.
Claims (5)
1. A double-network silk fibroin hydrogel is characterized in that,
the silk fibroin hydrogel is constructed by taking silk fibroin molecules as a matrix and introducing a host, an object and horseradish peroxidase into an amino acid side chain of the silk fibroin molecules;
the host is cyclodextrin, and the guest is adamantane;
the preparation method of the double-network silk fibroin hydrogel comprises the following steps:
(1) preparing carboxylated silk fibroin powder;
(2) preparing a silk fibroin-host solution;
(3) preparing a silk fibroin-guest solution;
(4) adding a horseradish peroxidase solution into the silk fibroin-main body solution, and uniformly mixing to obtain a first mixed solution;
(5) adding a hydrogen peroxide solution into the silk fibroin-object solution, and uniformly mixing to obtain a second mixed solution;
(6) uniformly mixing the first mixed solution and the second mixed solution, and standing to obtain the double-network silk fibroin hydrogel;
the step (2) comprises the following steps: dissolving the aminated main body in dimethyl sulfoxide solution to prepare aminated main body solution with concentration of 0.1-1000 mg/mL; dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with concentration of 0.1-1000 mg/mL; dissolving carboxylated silk fibroin powder in dimethyl sulfoxide solution to prepare carboxylated silk fibroin solution with the concentration of 0.05-10 mg/mL; mixing the aminated main body solution, an EDC.HCl solution and a carboxylated silk fibroin solution, reacting at room temperature, dialyzing, freezing and drying to obtain silk fibroin-main body powder; dissolving silk fibroin-main body powder in water to prepare a silk fibroin-main body solution with the mass fraction of 1-50%;
the step (3) comprises the following steps: preparing an object solution with the concentration of 0.1-1000 mg/mL; dissolving EDC.HCl in dimethyl sulfoxide solution to prepare EDC.HCl solution with concentration of 0.1-1000 mg/mL; dissolving carboxylated silk fibroin powder in dimethyl sulfoxide solution to prepare carboxylated silk fibroin solution with the concentration of 0.05-10 mg/mL; mixing the object solution, an EDC.HCl solution and a carboxylated silk fibroin solution, reacting at room temperature, dialyzing, freezing and drying to obtain silk fibroin-object powder; dissolving silk fibroin-object powder in water to prepare silk fibroin-object solution with the mass fraction of 1% -50%.
2. The method for preparing double-network silk fibroin hydrogel according to claim 1, characterized in that,
the step (1) comprises the following steps: degumming, dissolving and dialyzing silk to obtain a silk fibroin solution; chemically modifying the silk fibroin solution by chloroacetic acid to obtain a carboxylated silk fibroin solution with the concentration of 0.1-50%; and dialyzing the carboxylated silk fibroin solution, and freeze-drying to obtain carboxylated silk fibroin powder.
3. The method for preparing double-network silk fibroin hydrogel according to claim 1, characterized in that,
in the step (4), the content of horseradish peroxidase is 900-.
4. The method for preparing double-network silk fibroin hydrogel according to claim 1, characterized in that,
in the step (5), the concentration of the hydrogen peroxide solution is 160-170 mM.
5. The method for preparing double-network silk fibroin hydrogel according to claim 1, characterized in that,
in the step (6), the first mixed solution and the second mixed solution are uniformly mixed according to the molar ratio of the host to the guest of 100:1-1:100, and the mass fraction of the double-network silk fibroin hydrogel is 1% -50%.
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