CN109865161B - Spider silk protein bone nail and preparation method thereof - Google Patents
Spider silk protein bone nail and preparation method thereof Download PDFInfo
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- CN109865161B CN109865161B CN201711260874.0A CN201711260874A CN109865161B CN 109865161 B CN109865161 B CN 109865161B CN 201711260874 A CN201711260874 A CN 201711260874A CN 109865161 B CN109865161 B CN 109865161B
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- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Materials For Medical Uses (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention relates to the technical field of biomedicine. The invention provides a spider silk protein bone nail preparation method, which comprises the following steps: (1) preparing a spider silk protein solution; (2) and (3) injecting the spider silk protein solution into a mold, treating with methanol or ethanol, and drying to obtain the spider silk protein bone nails. The preparation method of the spider silk protein bone nail provided by the invention is simple to operate and can be industrially produced, the prepared spider silk protein bone nail has the advantages of high strength, high toughness, lightness, ultraviolet resistance, biodegradability and the like, and amino acids and polypeptides are formed after the spider silk protein bone nail is degraded by a human body, so that the spider silk protein bone nail has no toxic or side effect on the human body.
Description
Technical Field
The invention relates to the technical field of biomedicine, in particular to a spider silk protein bone nail and a preparation method thereof.
Background
Bone nails are common bone fixation instruments used in bone repair procedures. The traditional bone nail is made of metal materials such as stainless steel, titanium and alloy materials thereof, the materials have the advantages of high strength and good toughness, but secondary operation is carried out to take out the metal bone nail after a patient recovers, secondary pain is brought to the patient, and potential fracture danger is also left to the patient due to a cavity left after the secondary operation.
At present, degradable materials such as polylactic acid (PLA) and polyglycolide-lactide (PLGA) have been used in clinical bone repair. The bone nail made of absorbable polymer material can overcome many defects of the traditional bone nail material. However, materials such as polylactic acid (PLA) degrade to form acidic products, which negatively affects human tissues. Such materials, when degraded, cause acid accumulation and are not osteoconductive and thus do not promote the growth of bone cells. The material has high degradation speed and is not suitable for repairing main bearing bones. In addition, the mechanical properties of the material can not completely meet the clinical requirements of bone repair, and the mechanical properties of the material need to be improved by adding reinforcing fibers and the like.
Meanwhile, the PLA toughened ceramic-based material is used for preparing the bone built-in material, so that the biocompatibility is increased while the mechanical property of the bone nail is achieved, but the bone nail can not be completely degraded.
In addition, the bone nail made of the calcium phosphate bone material has degradability, good osteoconductivity and biocompatibility, and can bear compression load, but has low breaking strength, brittleness and easy fatigue.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, the present invention aims to provide a spider silk protein bone screw and a method for preparing the same, wherein the spider silk protein bone screw has the advantages of high strength, high toughness, lightness, ultraviolet resistance, biodegradability, etc., is amino acid and polypeptide after being degraded by a human body, and has no toxic effect on the human body.
The first method of the invention provides a spider silk protein bone nail preparation method, which comprises the following steps: (1) preparing a spider silk protein solution; (2) and (3) injecting the spider silk protein solution into a mold, treating with methanol or ethanol, and drying to obtain the spider silk protein bone nails.
In one embodiment of the present invention, in the step (2), after the methanol or ethanol treatment or drying, spider silk protein rods are obtained; and cutting the spider silk protein bar to obtain the spider silk protein bone nail.
In one embodiment of the invention, the spider silk protein solution is a solution of the spider silk protein hexafluoroisopropanol HFIP or an aqueous solution of the spider silk protein.
In one embodiment of the present invention, the concentration of spider silk protein in the spider silk protein solution is 0.01mg/mL to 300 mg/mL.
In one embodiment of the invention, said spider silk protein is selected from a natural spider silk protein or a genetically recombinant spider silk protein.
In one embodiment of the present invention, when said spider silk protein is a genetically recombinant spider silk protein, said step (1) comprises: placing engineering bacteria expressing gene recombinant spider silk protein in a Tris-HCl (PH 8)/NaCl/Imidazole solution with a first concentration, and performing thallus crushing treatment to obtain a first mixed solution; centrifuging the first mixed solution, and taking a supernatant to obtain a first supernatant; keeping the first supernatant in a constant-temperature water bath at 60-80 ℃ for a preset time, centrifuging, and taking the supernatant to obtain a second supernatant; adding the second supernatant into a nickel chromatographic column, removing impurity proteins by using a second concentration Tris-HCl (PH 8)/NaCl/Imidazole solution, and then eluting by using a third concentration Tris-HCl (PH 8)/NaCl/Imidazole solution to obtain a first initial solution of the gene recombinant spider silk; dialyzing the gene recombination spider silk protein eluent to obtain a second initial solution of the gene recombination spider silk; centrifuging the second initial solution of the gene recombinant spider silk, and removing supernatant to obtain a third initial solution of the gene recombinant spider silk protein; drying the third initial solution of the gene recombinant spider silk protein to obtain an anhydrous gene recombinant spider silk protein; dissolving the anhydrous gene recombinant spider silk protein in a solvent to obtain the spider silk protein solution.
In one embodiment of the present invention, the first concentration of Tris-HCl (PH 8)/NaCl/Imidazole solution has a Tris-HCl concentration range of 5-50mM, a NaCl concentration range of 50-200 mM, and an Imidazole concentration range of 1-10 mM. And in the second concentration Tris-HCl (PH 8)/NaCl/Imidazole solution, the concentration range of Tris-HCl is 5-50mM, the concentration range of NaCl is 50-200 mM, and the concentration range of Imidazole is 40-60 mM. In the third concentration Tris-HCl (PH 8)/NaCl/Imidazole solution, the concentration range of Tris-HCl is 5-50mM, the concentration range of NaCl is 50-200 mM, and the concentration range of Imidazole is 200-.
In one embodiment of the invention, the step of drying the third initial solution of the genetically recombinant spider silk protein comprises: freezing the third initial solution of the gene recombinant spider silk protein in an environment of-80-0 ℃; freezing in a freeze dryer for 1-500 hours in vacuum; wherein the vacuum pressure of the freeze dryer is 0.001-1 mBar, and the temperature is-80-0 ℃.
In one embodiment of the present invention, the step (1) comprises: dissolving 1 weight part of anhydrous spider silk protein in 0.5-10 weight parts of hexafluoroisopropanol to obtain a spider silk protein solution; wherein the temperature of the anhydrous spider silk protein and the hexafluoroisopropanol is 5-50 ℃.
In one embodiment of the present invention, the step (1) comprises: dissolving 1 part by weight of anhydrous spider silk protein in 01-10 parts by weight of water to obtain a spider silk protein solution; wherein the temperature of the anhydrous spider silk protein and the water is 5-50 ℃.
In one embodiment of the present invention, in the step (2), the methanol treatment comprises: immersing the mold injected with the spider silk protein solution in liquid methanol, and replacing the methanol solution every 1-50 hours for 1-50 times to obtain a spider silk protein egg solid; and taking out the spider silk protein solid from the mold, continuously soaking the spider silk protein solid by using methanol, and replacing the methanol solution every 1-50 hours, wherein the replacement times are 1-50 times.
In one embodiment of the present invention, in the step (2), the ethanol treatment comprises: immersing the die filled with the spider silk protein solution into liquid ethanol, and replacing the ethanol solution every 1-50 hours for 1-50 times to obtain a spider silk protein egg solid; and taking the spider silk protein solid out of the mold, continuously soaking the spider silk protein solid with ethanol, and replacing the ethanol solution every 1-50 hours, wherein the replacement times are 1-50 times.
In one embodiment of the present invention, in the step (2), the drying includes: and volatilizing methanol or ethanol in the spider silk protein and air-drying in a ventilation environment, wherein the temperature of the ventilation environment is 10-50 ℃.
In an embodiment of the present invention, the method further includes: (3) soaking the spider silk protein bone nails obtained in the step (2) in an active drug solution to obtain spider silk protein bone nails containing the active drug; wherein, the concentration of the medicine in the active medicine solution is 0.001 mu g/mL-5 mg/mL, and the soaking time is 1 minute-500 hours.
In one embodiment of the invention, the active agent comprises an active factor that promotes bone growth; wherein the active factors promoting bone growth comprise any one or more of the following: BMP, FGF, PDGF, VEGF, TGF, IGF.
In one embodiment of the present invention, the active agent comprises an antibacterial and anti-inflammatory agent; wherein, the antibacterial and anti-inflammatory medicine comprises any one or more of the following medicines: penicillins, cephalosporins, aminoglycosides, macrolides, tetracyclines, quinolones, sulfonamides.
In one embodiment of the present invention, the step (1) comprises: adding 0.5 weight part of antibacterial and anti-inflammatory drugs and/or active factors for promoting bone growth into 0.5-10 weight parts of solvent to obtain a drug solvent; adding 1 part by weight of a spider silk protein to the drug solvent to obtain the spider silk protein solution; wherein, the antibacterial and anti-inflammatory medicine comprises any one or more of the following medicines: penicillins, cephalosporins, aminoglycosides, macrolides, tetracyclines, quinolones, sulfonamides; the active factors promoting bone growth comprise any one or more of the following: BMP, FGF, PDGF, VEGF, TGF, IGF.
In one embodiment of the present invention, in the step (1), the spider silk protein is prepared into an anhydrous spider silk protein, and the obtained anhydrous spider silk protein is dissolved in a solvent to obtain the spider silk protein solution.
In one embodiment of the present invention, in the step (1), an aqueous solution of spider silk protein is prepared, and then freeze-dried to obtain an anhydrous spider silk protein, and the obtained anhydrous spider silk protein is dissolved in a solvent to obtain the spider silk protein solution.
The invention also provides a spider silk protein bone nail, which is prepared by the spider silk protein bone nail preparation method of the first aspect.
A third aspect of the invention provides the use of the aforementioned spider silk protein bone nails in the preparation of bone fixation instruments.
Compared with the prior art, the invention has the following beneficial effects:
the spider silk protein bone nail prepared by the preparation method of the invention has good affinity with human body, no immunological rejection reaction, good compatibility in human tissue and no toxic action on human body; according to the preparation method of the spider silk protein bone nail provided by the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so as to meet different clinical requirements.
Drawings
FIG. 1 is a schematic flow chart of a spider silk protein bone nail preparation method provided by an embodiment of the invention.
FIG. 2 is a schematic structural diagram of a spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment of the invention.
FIG. 3 is a relationship between the soaking time of a spider silk protein bone nail in an active drug solution and the loading depth of an active drug in the method for preparing a spider silk protein bone nail according to an embodiment of the present invention.
FIG. 4 is a mechanical property curve of a spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment of the invention.
FIG. 5 is a diagram showing the inhibitory effect of the spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment of the present invention on bacteria.
Detailed Description
Before the present embodiments are further described, it is to be understood that the scope of the invention is not limited to the particular embodiments described below; it is also to be understood that the terminology used in the examples is for the purpose of describing particular embodiments, and is not intended to limit the scope of the present invention; in the description and claims of the present application, the singular forms "a", "an" and "the" include plural referents unless the context clearly dictates otherwise.
When numerical ranges are given in the examples, it is understood that both endpoints of each of the numerical ranges and any value therebetween can be selected unless the invention otherwise indicated. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In addition to the specific methods, devices, and materials used in the examples, any methods, devices, and materials similar or equivalent to those described in the examples may be used in the practice of the invention in addition to the specific methods, devices, and materials used in the examples, in keeping with the knowledge of one skilled in the art and with the description of the invention.
Example 1
The embodiment 1 of the invention provides a spider silk protein bone nail preparation method, which comprises the following steps as shown in fig. 1.
The spider silk protein solution can be a solution of natural spider silk protein or a solution of gene recombinant spider silk protein. Next, genetic recombination spider silk proteins will be described as an example. Engineering bacteria which are subjected to high-density fermentation and express the gene recombinant spider silk protein are dissolved in a first Tris-HCl (PH 8)/NaCl/Imidazole solution (Tris-HCl concentration is 20mM, NaCl concentration is 150mM, and Imidazole range is 5mM), and the bacteria are crushed by a bacteria breaker. Centrifuging the disrupted bacteria, and collecting the supernatant; then preserving the heat of the supernatant for 24 hours in a constant-temperature water bath at 80 ℃, centrifuging and collecting the supernatant; adding the supernatant into a nickel chromatographic column, washing off the hybrid protein by using a second Tris-HCl (PH 8)/NaCl/Imidazole solution (Tris-HCl concentration is 20mM, NaCl concentration is 150mM, and Imidazole range is 50mM), and eluting the target gene recombinant spider silk protein by using a third Tris-HCl (PH 8)/NaCl/Imidazole solution (Tris-HCl concentration is 20mM, NaCl concentration is 150mM, and Imidazole range is 250mM) and collecting; putting the collected gene recombinant spider silk protein solution into a dialysis bag, placing the dialysis bag in a phosphate buffer solution for 24 hours of gradient dialysis, and transferring the dialysis bag into ultrapure water for 48 hours of dialysis; and (3) centrifugally separating the dialyzed gene recombination spider silk protein solution, and collecting supernatant to obtain the required gene recombination spider silk protein initial solution.
The gene recombinant spider silk protein initial solution can be dried to obtain anhydrous gene recombinant spider silk protein, and then the anhydrous gene recombinant protein is dissolved in a solvent to obtain spider silk protein solution. The initial solution of the gene recombinant spider silk protein can also be concentrated to obtain the spider silk protein solution.
The drying of the gene recombinant spider silk protein initial solution can be freeze drying, specifically, the gene recombinant spider silk protein initial solution is firstly frozen for 12 hours in an environment of-80 ℃, and then is subjected to vacuum freezing treatment for 100 hours in a freeze dryer, wherein the vacuum pressure of the freeze dryer is 0.02mBar, and the temperature is-20 ℃.
The spider silk protein solution can be a gene recombinant spider silk protein Hexafluoroisopropanol (HFIP) solution. The anhydrous gene recombinant protein can be dissolved in Hexafluoroisopropanol (HFIP) to obtain the hexafluoroisopropanol solution of the gene recombinant spider silk protein. Specifically, when the anhydrous gene recombinant spider silk protein is dissolved, the mass ratio of the anhydrous gene recombinant spider silk protein to hexafluoroisopropanol is 1:4, the temperature is 25 ℃, and the dissolving time is 48 hours. Meanwhile, a low-concentration anti-inflammatory drug ampicillin is added in advance, and the mass ratio of ampicillin to anhydrous gene recombinant spider silk protein is 0.5mg to 1 g.
The spider silk protein solution can be a gene recombinant spider silk protein aqueous solution. Preparing the spider silk protein solution by dissolving 1 part by weight of anhydrous spider silk protein in 1 part by weight of water to obtain the spider silk protein solution; wherein the temperature of the anhydrous spider silk protein and the water is 5 ℃.
And 2, injecting the spider silk protein solution into a mold, and carrying out methanol treatment and drying to obtain the spider silk protein bone nails.
The mold can be a bone nail mold, and the spider silk protein bone nail can be directly prepared by the bone nail mold; the die can also be a cylindrical die, and a spider silk protein bar can be prepared firstly and then cut into the spider silk protein bone nails.
Next, step 2 will be specifically described by taking a cylindrical mold and a genetic recombinant spider silk as an example. The die was a cylindrical die (having a through hole of 100 μm diameter on the surface) of 10mm in diameter. And (3) injecting the spider silk protein solution into a mold, and then placing the mold in a methanol environment for treatment. The methanol treatment process comprises the following steps: immersing a die containing a spider silk protein solution into liquid methanol, and replacing the methanol solution once every 12 hours, wherein the liquid replacement times are 10 times, so as to obtain a gene recombinant spider silk protein solid; then taking out the gene recombinant spider silk protein solid from the mould, continuously soaking the solid with methanol, and replacing the methanol solution every 24 hours for 5 times.
Cutting the gene recombinant spider silk protein bar, namely the gene recombinant spider silk protein bone nail into a bone nail shape. Specifically, the gene recombinant spider silk protein rods can be cut into bone nails according to specific clinical requirements for bone repair, as shown in fig. 2.
Soaking the spider silk protein bone nail in an aqueous solution of an active drug, wherein the surface of the bone nail contains the drug with the effects of promoting bone growth and/or resisting bacteria and diminishing inflammation within a certain depth. Specifically, the water solution of the active drug is ampicillin solution, the concentration is 5mg/mL, the soaking time is 24 hours, and the drug loading depth is about 100 micrometers. The relationship between the soaking time and the depth of drug loading is shown in fig. 3.
And (5) airing the spider silk protein bone nails in a ventilation environment, sterilizing and packaging. Specifically, the air is dried in a fume hood at 24 ℃ (room temperature) for 12 hours, and the sterilization method is Co60 radioactive sterilization method.
The spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment 1 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so as to meet different clinical requirements.
The mechanical properties of the spider silk protein bone nail provided by the embodiment 1 of the invention are tested, and the result is shown in fig. 4, the elastic modulus value of the spider silk protein bone nail is 3.5GPa, which is closer to the elastic modulus of human skeleton of 4-6 GPa, so that the spider silk protein bone nail does not cause stress shielding effect in the bone implantation and healing processes, and the defects of metal materials in the aspect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drug provided by the embodiment of the invention; wherein the concentration of the ampicillin drug water solution is 5mg/mL, and the soaking time of the spider silk protein bone nails therein is 24 hours; after wiping off the residual drug solution on the surface of the bone nail, the bone nail is dried in a ventilated environment for 12 hours, and then inserted and cultured on an escherichia coli gel plate for 24 hours. As shown in FIG. 5, it can be found that the bone screw loaded with ampicillin drug has good bacteria inhibition effect on the E.coli gel plate.
The spider silk protein bone nail provided by the embodiment 1 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is loaded inside or on the surface of the medicine box, so that the medicine treatment function is realized.
Example 2
In step 1, the first Tris-HCl (PH 8)/NaCl/Imidazole solution is a solution with a Tris-HCl concentration of 5mM, a NaCl concentration of 200mM, an Imidazole range of 10 mM); the second Tris-HCl (PH 8)/NaCl/Imidazole solution was a Tris-HCl 5mM concentration, a NaCl 200mM concentration, and an Imidazole range 40mM solution; the third Tris-HCl (PH 8)/NaCl/Imidazole solution was a solution with a Tris-HCl concentration of 50mM, a NaCl concentration of 200mM, and an Imidazole range of 300 mM. The other operations were the same as in example 1.
The spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment 2 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so as to meet different clinical requirements.
The mechanical properties of the spider silk protein bone nail provided by the embodiment 2 of the invention are tested, the elastic modulus value is about 3.5GPa, the elastic modulus value is closer to the elastic modulus of human skeleton of 4-6 GPa, the stress shielding effect cannot be caused in the bone implantation and healing processes, and the defects of metal materials in this respect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drug provided by the embodiment of the invention; as a result, the bone nail carrying the ampicillin drug has good bacteria inhibition effect on the Escherichia coli gel plate.
The spider silk protein bone nail provided by the embodiment 2 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is loaded inside or on the surface of the medicine box, so that the medicine treatment function is realized.
In the case of the embodiment 3, the following examples,
in step 1, dissolving natural spider silk protein in water, freezing at-40 deg.C, and vacuum freezing in vacuum freeze dryer for 250 hr under vacuum pressure of 0.001mBar at-80 deg.C; obtaining anhydrous natural spider silk protein, and dissolving 1 weight part of the anhydrous natural spider silk protein in 0.5 weight part of hexafluoroisopropanol at 27.5 ℃ to obtain the spider silk protein solution. The other steps are the same as in example 1.
The spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment 3 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so as to meet different clinical requirements.
The mechanical properties of the spider silk protein bone nail provided by the embodiment 3 of the invention are tested, the elastic modulus value is about 3.5GPa, the elastic modulus value is closer to the elastic modulus of human skeleton of 4-6 GPa, the stress shielding effect cannot be caused in the bone implantation and healing processes, and the defects of metal materials in this respect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drug provided by the embodiment of the invention; as a result, the bone nail carrying the ampicillin drug has good bacteria inhibition effect on the Escherichia coli gel plate.
The spider silk protein bone nail provided by the embodiment 3 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is loaded inside or on the surface of the medicine box, so that the medicine treatment function is realized.
Example 4
In the step 1, dissolving spider silk protein in water, freezing at 0 ℃, and carrying out vacuum freezing treatment in a vacuum freeze dryer for 500 hours at the vacuum pressure of 1mBar and the temperature of-40 ℃; obtaining anhydrous spider silk protein, dissolving 1 weight part of anhydrous natural spider silk protein in 5 weight parts of hexafluoroisopropanol at 50 ℃ to obtain the spider silk protein solution. The other steps are the same as in example 1.
The spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment 4 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so as to meet different clinical requirements.
The mechanical properties of the spider silk protein bone nail provided by the embodiment 4 of the invention are tested, the elastic modulus value is about 3.5GPa, the elastic modulus value is closer to the elastic modulus of human skeleton 4-6 GPa, the stress shielding effect cannot be caused in the bone implantation and healing processes, and the defects of metal materials in this respect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drug provided by the embodiment of the invention; as a result, the bone nail carrying the ampicillin drug has good bacteria inhibition effect on the Escherichia coli gel plate.
The spider silk protein bone nail provided by the embodiment 4 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is loaded inside or on the surface of the medicine box, so that the medicine treatment function is realized.
Example 5
In step 1, 1 part by weight of the freeze-dried anhydrous spider silk protein is dissolved in 10 parts by weight of hexafluoroisopropanol at a dissolution temperature of 5 ℃ to obtain the spider silk protein solution. The other steps are the same as in example 1.
The spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment 5 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so as to meet different clinical requirements.
The spider silk protein bone nail provided by the embodiment 5 of the invention is tested for mechanical properties, the elastic modulus value is about 3.5GPa, and is closer to the elastic modulus of human skeleton of 4-6 GPa, so that the stress shielding effect is not caused in the bone implantation and healing processes, and the defects of metal materials in this respect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drug provided by the embodiment of the invention; as a result, the bone nail carrying the ampicillin drug has good bacteria inhibition effect on the Escherichia coli gel plate.
The spider silk protein bone nail provided by the embodiment 5 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is loaded inside or on the surface of the medicine box, so that the medicine treatment function is realized.
Example 6
In step 1, 1 part by weight of the freeze-dried anhydrous spider silk protein was dissolved in 5.5 parts by weight of water at a dissolution temperature of 27.5 ℃ to obtain a spider silk protein solution, and the other operations were the same as in example 1.
The spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment 6 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so as to meet different clinical requirements.
The spider silk protein bone nail provided by the embodiment 6 of the invention is tested for mechanical properties, the elastic modulus value is about 3.5GPa, and is closer to the elastic modulus of human skeleton of 4-6 GPa, so that the stress shielding effect is not caused in the bone implantation and healing processes, and the defects of metal materials in this respect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drug provided by the embodiment of the invention; as a result, the bone nail carrying the ampicillin drug has good bacteria inhibition effect on the Escherichia coli gel plate.
The spider silk protein bone nail provided by the embodiment 6 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is loaded inside or on the surface of the medicine box, so that the medicine treatment function is realized.
Example 7
In step 1, 1 part by weight of the freeze-dried anhydrous spider silk protein was dissolved in 10 parts by weight of water at 50 ℃ to obtain a spider silk protein solution, and the other operations were the same as in example 1.
The spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment 7 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so as to meet different clinical requirements.
The mechanical property of the spider silk protein bone nail provided by the embodiment 7 of the invention is tested, the elastic modulus value is about 3.5GPa, and is closer to the elastic modulus of human skeleton of 4-6 GPa, so that the stress shielding effect can not be caused in the process of bone implantation and healing, and the defects of metal materials in this respect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drug provided by the embodiment of the invention; as a result, the bone nail carrying the ampicillin drug has good bacteria inhibition effect on the Escherichia coli gel plate.
The spider silk protein bone nail provided by the embodiment 7 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is loaded inside or on the surface of the medicine box, so that the medicine treatment function is realized.
Example 8
In step 2, ethanol was used instead of methanol, and the other operations were the same as in example 1.
The spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment 8 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so as to meet different clinical requirements.
The spider silk protein bone nail provided by the embodiment 8 of the invention is tested for mechanical properties, the elastic modulus value is about 3.5GPa, and is closer to the elastic modulus of human skeleton of 4-6 GPa, so that the stress shielding effect is not caused in the bone implantation and healing processes, and the defects of metal materials in this respect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drug provided by the embodiment of the invention; as a result, the bone nail carrying the ampicillin drug has good bacteria inhibition effect on the Escherichia coli gel plate.
The spider silk protein bone nail provided by the embodiment 8 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is loaded inside or on the surface of the medicine box, so that the medicine treatment function is realized.
Example 9
In step 2, the spider silk protein solution is injected into a mold and then placed in a methanol environment for treatment. The methanol treatment process comprises the following steps: immersing a die containing a spider silk protein solution into liquid methanol, and replacing the methanol solution every 1 hour for 26 times to obtain a gene recombinant spider silk protein solid; then taking out the gene recombinant spider silk protein solid from the mould, continuously soaking the solid with methanol, and replacing the methanol solution every 50 hours for 1 time. The other operations were the same as in example 1.
The spider silk protein bone nail prepared by the method for preparing the spider silk protein bone nail provided by the embodiment 9 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and the degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so that different clinical requirements can be met.
The spider silk protein bone nail provided by the embodiment 9 of the invention is tested for mechanical properties, the elastic modulus value is about 3.5GPa, and is closer to the elastic modulus of human skeleton of 4-6 GPa, so that the stress shielding effect is not caused in the bone implantation and healing processes, and the defects of metal materials in this respect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drug provided by the embodiment of the invention; as a result, the bone nail carrying the ampicillin drug has good bacteria inhibition effect on the Escherichia coli gel plate.
The spider silk protein bone nail provided by the embodiment 9 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is loaded inside or on the surface of the medicine box, so that the medicine treatment function is realized.
Example 10
In step 2, the spider silk protein solution is injected into a mold and then placed in a methanol environment for treatment. The methanol treatment process comprises the following steps: immersing a die containing a spider silk protein solution into liquid methanol, and replacing the methanol solution once every 50 hours, wherein the liquid replacement times are 50 times, so as to obtain a gene recombinant spider silk protein solid; then taking out the gene recombinant spider silk protein solid from the mould, continuously soaking the solid with methanol, and replacing the methanol solution every 1 hour for 50 times. The other operations were the same as in example 1.
The spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment 10 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so as to meet different clinical requirements.
The spider silk protein bone nail provided by the embodiment 10 of the invention is tested for mechanical properties, the elastic modulus value is about 3.5GPa, and is closer to the elastic modulus of human skeleton of 4-6 GPa, so that the stress shielding effect is not caused in the bone implantation and healing processes, and the defects of metal materials in this respect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drug provided by the embodiment of the invention; as a result, the bone nail carrying the ampicillin drug has good bacteria inhibition effect on the Escherichia coli gel plate.
The spider silk protein bone nail provided by the embodiment 10 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is loaded inside or on the surface of the medicine box, so that the medicine treatment function is realized.
Example 11
In step 2, the ventilated ambient temperature is 50 deg.f. The other operations were the same as in example 1.
The spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment 11 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and the degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so that different clinical requirements can be met.
The mechanical property of the spider silk protein bone nail provided by the embodiment 11 of the invention is tested, the elastic modulus value is about 3.5GPa, and is closer to the elastic modulus of human skeleton of 4-6 GPa, so that the stress shielding effect can not be caused in the process of bone implantation and healing, and the defects of metal materials in this respect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drug provided by the embodiment of the invention; as a result, the bone nail carrying the ampicillin drug has good bacteria inhibition effect on the Escherichia coli gel plate.
The spider silk protein bone nail provided by the embodiment 11 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is loaded inside or on the surface of the medicine box, so that the medicine treatment function is realized.
Example 12
In step 2, the temperature of the ventilated environment is 10 deg.f. The other operations were the same as in example 1.
The spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment 12 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so as to meet different clinical requirements.
The spider silk protein bone nail provided by the embodiment 12 of the invention is tested for mechanical properties, the elastic modulus value is about 3.5GPa, and is closer to the elastic modulus of human skeleton of 4-6 GPa, so that the stress shielding effect is not caused in the bone implantation and healing processes, and the defects of metal materials in this respect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drug provided by the embodiment of the invention; as a result, the bone nail carrying the ampicillin drug has good bacteria inhibition effect on the Escherichia coli gel plate.
The spider silk protein bone nail provided by the embodiment 12 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is loaded inside or on the surface of the medicine box, so that the medicine treatment function is realized.
Example 13
The same procedure as in example 1 was repeated except that the aqueous solution of the active drug was a penicillin solution at a concentration of 1mg/mL and the soaking time was 50 hours.
The spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment 13 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so as to meet different clinical requirements.
The mechanical properties of the spider silk protein bone nail provided by the embodiment 13 of the invention are tested, the elastic modulus value is about 3.5GPa, the elastic modulus value is closer to the elastic modulus of human skeleton of 4-6 GPa, the stress shielding effect cannot be caused in the bone implantation and healing processes, and the defects of metal materials in this respect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drug provided by the embodiment of the invention; as a result, the bone nail carrying the ampicillin drug has good bacteria inhibition effect on the Escherichia coli gel plate.
The spider silk protein bone nail provided by the embodiment 13 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is carried in the inner part or on the surface of the medicine box, so that the medicine treatment function is realized.
Example 14
The same procedure as in example 1 was repeated except that the aqueous solution of the active drug was ampicillin at a concentration of 0.001. mu.g/mL and the soaking time was 500 hours.
The spider silk protein bone nail prepared by the spider silk protein bone nail preparation method provided by the embodiment 14 of the invention has good affinity with a human body, no immune rejection reaction, good compatibility in human tissues and no toxic action on the human body; according to the preparation method of the spider silk protein bone nail provided by the embodiment of the invention, a functional drug is added at the dissolving stage of the spider silk protein, or the spider silk protein bone nail is soaked in a drug water solution, so that drugs can be loaded inside and on the surface of the bone nail, and a drug treatment function can be realized. In addition, the mechanical property and degradation speed of the bone nail can be realized through parameters such as the molecular weight, the mechanical gene group, the crystallinity and the like of the spider silk protein, so as to meet different clinical requirements.
The spider silk protein bone nail provided by the embodiment 14 of the invention is tested for mechanical properties, the elastic modulus value is about 3.5GPa, and is closer to the elastic modulus of human skeleton of 4-6 GPa, so that the stress shielding effect is not caused in the bone implantation and healing processes, and the defects of metal materials in this respect are overcome.
Carrying out a bacteria inhibition effect experiment on the spider silk protein bone nail loaded with the ampicillin drugs provided by the embodiment of the invention; as a result, the bone nail carrying the ampicillin drug has good bacteria inhibition effect on the Escherichia coli gel plate.
The spider silk protein bone nail provided by the embodiment 14 of the invention has good affinity with a human body, no immune rejection, good compatibility in human tissues and no toxic action on the human body; the mechanical property of the composite material is close to that of human skeleton, so that the stress shielding effect in the process of bone implantation and healing is avoided; the medicine is loaded inside or on the surface of the medicine box, so that the medicine treatment function is realized.
In conclusion, the present invention effectively overcomes various disadvantages of the prior art and has high industrial utilization value.
The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Claims (8)
1. A method for preparing a spider silk protein bone nail is characterized by comprising the following steps:
(1) preparing a spider silk protein solution;
the spider silk protein is a gene recombinant spider silk protein, and the step (1) comprises the following steps:
placing engineering bacteria expressing gene recombinant spider silk protein in a Tris-HCl (PH 8)/NaCl/Imidazole solution with a first concentration, and performing thallus crushing treatment to obtain a first mixed solution; in the first concentration Tris-HCl (PH 8)/NaCl/Imidazole solution, the concentration range of Tris-HCl is 5-50mM, the concentration range of NaCl is 50-200 mM, and the concentration range of Imidazole is 1-10 mM;
centrifuging the first mixed solution, and taking a supernatant to obtain a first supernatant;
keeping the first supernatant in a constant-temperature water bath at 60-80 ℃ for a preset time, centrifuging, and taking the supernatant to obtain a second supernatant;
adding the second supernatant into a nickel chromatographic column, removing impurity proteins by using a second concentration Tris-HCl (PH 8)/NaCl/Imidazole solution, and then eluting by using a third concentration Tris-HCl (PH 8)/NaCl/Imidazole solution to obtain a first initial solution of the gene recombinant spider silk; in the second concentration Tris-HCl (PH 8)/NaCl/Imidazole solution, the concentration range of Tris-HCl is 5-50mM, the concentration range of NaCl is 50-200 mM, and the concentration range of Imidazole is 40-60 mM; in the third concentration Tris-HCl (PH 8)/NaCl/Imidazole solution, the concentration range of Tris-HCl is 5-50mM, the concentration range of NaCl is 50-200 mM, and the concentration range of Imidazole is 200-;
dialyzing the gene recombination spider silk protein eluent to obtain a second initial solution of the gene recombination spider silk;
centrifuging the second initial solution of the gene recombinant spider silk, and removing supernatant to obtain a third initial solution of the gene recombinant spider silk protein;
drying the third initial solution of the gene recombinant spider silk protein to obtain an anhydrous gene recombinant spider silk protein; the step of drying the third initial solution of the gene recombinant spider silk protein comprises the following steps: freezing the third initial solution of the gene recombinant spider silk protein in an environment of-80-0 ℃; vacuum freezing in a freeze dryer for 1-500 hours; wherein the vacuum pressure of the freeze dryer is 0.001-1 mBar, and the temperature is-80-0 ℃;
dissolving the anhydrous genetic recombinant spider silk protein in a solvent to obtain the spider silk protein solution; the spider silk protein solution is a spider silk protein hexafluoroisopropanol HFIP solution or a spider silk protein water solution;
the step (1) comprises the following steps: dissolving 1 weight part of anhydrous gene recombinant spider silk protein in 0.5-10 weight parts of hexafluoroisopropanol or 1-10 weight parts of water to obtain a spider silk protein solution; wherein the temperature of the anhydrous genetic recombination spider silk protein and the hexafluoroisopropanol is 5-50 ℃, and the temperature of the anhydrous genetic recombination spider silk protein and the water is 5-50 ℃;
(2) injecting the spider silk protein solution into a mold, treating with methanol or ethanol, and drying to obtain spider silk protein bone nails;
in the step (2), after the spider silk protein bar is treated by the methanol or the ethanol and dried, the spider silk protein bar is obtained; and cutting the spider silk protein bar to obtain the spider silk protein bone nail.
2. The spider silk protein bone nail preparation method according to claim 1, characterized in that in step (2), the methanol treatment comprises: immersing the die injected with the spider silk protein solution into liquid methanol, and replacing the methanol solution every 1-50 hours for 1-50 times to obtain a spider silk protein solid; taking out the spider silk protein solid from the mold, continuously soaking the spider silk protein solid by using methanol, and replacing the methanol solution every 1-50 hours, wherein the replacement times are 1-50 times;
in the step (2), the ethanol treatment comprises: immersing the die filled with the spider silk protein solution into liquid ethanol, and replacing the ethanol solution every 1-50 hours for 1-50 times to obtain a spider silk protein solid; and taking out the spider silk protein solid from the mold, continuously soaking the spider silk protein solid by using ethanol, and replacing the ethanol solution every 1-50 hours, wherein the replacement times are 1-50 times.
3. The spider silk protein bone nail production method according to claim 1, characterized in that in step (2), the drying comprises: and volatilizing methanol or ethanol in the spider silk protein and air-drying in a ventilation environment, wherein the temperature of the ventilation environment is 10-50 ℃.
4. The spider silk protein bone nail production method according to claim 1, further comprising: (3) soaking the spider silk protein bone nails obtained in the step (2) in an active drug solution to obtain spider silk protein bone nails containing the active drug; wherein, the concentration of the medicine in the active medicine solution is 0.001 mu g/mL-5 mg/mL, and the soaking time is 1 minute-500 hours.
5. The spider silk protein bone nail production method according to claim 4, characterized in that the active drug comprises an active factor that promotes bone growth; wherein the active factors promoting bone growth comprise any one or more of the following: BMP, FGF, PDGF, VEGF, TGF, IGF.
6. The spider silk protein bone screw production method according to claim 4, characterized in that the active drug comprises an antibacterial anti-inflammatory drug; wherein, the antibacterial and anti-inflammatory medicine comprises any one or more of the following medicines: penicillins, cephalosporins, aminoglycosides, macrolides, tetracyclines, quinolones, sulfonamides.
7. A spider silk protein bone nail, characterized by being produced by the spider silk protein bone nail production method according to any one of claims 1 to 6.
8. Use of a spider silk protein bone peg of any one of claims 1 to 6 or of claim 7 for the manufacture of a bone fixation device.
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| CN201711260874.0A CN109865161B (en) | 2017-12-04 | 2017-12-04 | Spider silk protein bone nail and preparation method thereof |
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| CN201711260874.0A CN109865161B (en) | 2017-12-04 | 2017-12-04 | Spider silk protein bone nail and preparation method thereof |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101291694A (en) * | 2005-08-17 | 2008-10-22 | 牛津生物材料有限公司 | Implantable cartilage tissue repair device |
| CN102532295A (en) * | 2004-07-22 | 2012-07-04 | Am丝绸有限责任公司 | recombinant spider silk protein |
| CN106668956A (en) * | 2015-11-11 | 2017-05-17 | 中国科学院上海微系统与信息技术研究所 | Fibroin bone nail and preparation method thereof |
| CN107041972A (en) * | 2016-02-05 | 2017-08-15 | 上海沐研斯生物科技有限公司 | A kind of fibroin nail and preparation method thereof |
| CN107213521A (en) * | 2017-05-31 | 2017-09-29 | 合肥创沃科技有限公司 | A kind of preparation method of spider silk composite bioactivity glass tissue renovation material |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102532295A (en) * | 2004-07-22 | 2012-07-04 | Am丝绸有限责任公司 | recombinant spider silk protein |
| CN101291694A (en) * | 2005-08-17 | 2008-10-22 | 牛津生物材料有限公司 | Implantable cartilage tissue repair device |
| CN106668956A (en) * | 2015-11-11 | 2017-05-17 | 中国科学院上海微系统与信息技术研究所 | Fibroin bone nail and preparation method thereof |
| CN107041972A (en) * | 2016-02-05 | 2017-08-15 | 上海沐研斯生物科技有限公司 | A kind of fibroin nail and preparation method thereof |
| CN107213521A (en) * | 2017-05-31 | 2017-09-29 | 合肥创沃科技有限公司 | A kind of preparation method of spider silk composite bioactivity glass tissue renovation material |
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