CN109867811B - Porous-solid composite silk material and preparation method thereof - Google Patents
Porous-solid composite silk material and preparation method thereof Download PDFInfo
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Abstract
The invention provides a preparation method of a porous-solid composite silk material, which comprises the following steps: (1) dissolving silk fibroin in hexafluoroisopropanol to obtain a silk fibroin solution; (2) injecting the silk fibroin solution into a porous mold, and standing; wherein the porous mold comprises a peripheral wall and a protective shell, the peripheral wall having a plurality of openings; (3) filling water-soluble salt particles into the silk fibroin solution in the porous mold to obtain a water-soluble salt particle-silk fibroin solution mixture; (4) soaking the porous mold containing the water-soluble salt particle-silk fibroin solution mixture after the protective shell is removed in methanol to prepare silk fibroin solid containing water-soluble salt particles; (5) and removing methanol and water-soluble salt particles in the silk fibroin solid containing the water-soluble salt particles to obtain the porous-solid composite silk fibroin material.
Description
Technical Field
The invention relates to the technical field of organic high polymer materials, in particular to a porous-solid composite silk material and a preparation method thereof.
Background
The biobased material with excellent performance, environmental protection, various varieties, high added value and wide application is one of the important fields of strategic emerging industry and biomass industry development in China. Biomacromolecule materials, especially natural products, have many unique advantages in the field of polymer materials, such as good biocompatibility, degradability, and wide source.
As a biological material with a long application history, the fibroin material is one of natural protein materials which are utilized and developed by people at first. Wherein, silk fibroin is the main constituent material of silk (accounting for about 70 percent of the mass of the silk) and is a novel green bio-based material with modern technological characteristics. Compared with other protein materials and organic polymer materials, silk fibroin has unique and excellent biological, optical and mechanical properties, including high light transmittance, no human rejection, human body absorbability, controllable dissolution and degradation rates, high mechanical strength, wide sources and the like, thereby playing an important role in a plurality of new leading-edge fields.
At present, the silk fibroin block is mainly in the form of a porous scaffold, and a porous-solid composite silk fibroin material is not available, so that the silk fibroin block is limited to have wide application in the fields of biological medicine, novel microelectronics and the like. Therefore, how to prepare the porous-solid composite silk fibroin material is the subject of considerable research.
Disclosure of Invention
In view of the above-mentioned drawbacks of the prior art, the present invention provides a porous-solid composite silk material and a preparation method thereof, so as to prepare a porous-solid composite silk material with adjustable porosity and void diameter, firm combination of porous structure and solid structure, and controllable biodegradation and mechanical properties, which can be widely applied in the field of bio-implant medical treatment.
The first method of the invention provides a preparation method of a porous-solid composite silk material, which comprises the following steps: (1) dissolving the silk fibroin in hexafluoroisopropanol to obtain a silk fibroin solution; (2) injecting the silk fibroin solution into a porous mold, and standing; wherein the porous mold comprises a peripheral wall and a protective shell, the peripheral wall having a plurality of openings; (3) filling water-soluble salt particles into the silk fibroin solution in the porous mold to obtain a water-soluble salt particle-silk fibroin solution mixture; (4) soaking the porous mold containing the water-soluble salt particle-silk fibroin solution mixture after the protective shell is removed in methanol to prepare silk fibroin solid containing water-soluble salt particles; (5) and removing methanol and water-soluble salt particles in the silk fibroin solid containing the water-soluble salt particles to obtain the porous-solid composite silk fibroin material.
In one embodiment of the invention, in step (1), the silk fibroin is anhydrous silk fibroin.
In one embodiment of the invention, before dissolving the silk fibroin in hexafluoroisopropanol, preparing a silk fibroin aqueous solution, and freeze-drying the silk fibroin aqueous solution to obtain anhydrous silk fibroin dry powder; and dissolving the anhydrous silk fibroin dry powder in hexafluoroisopropanol to obtain the silk fibroin solution.
In one embodiment of the present invention, the step (1) comprises: placing the silkworm cocoon in a sodium carbonate solution, and heating to form silk; first drying; adding LiBr or LiSCN solution, and preserving heat for preset time to obtain mixed solution; dialyzing the mixed solution, centrifuging the dialyzed solution, and taking supernatant to obtain a silk fibroin aqueous solution; and secondly, freeze-drying the silk fibroin solution to obtain the silk fibroin.
Preferably, the silkworm cocoon is a silkworm cocoon shell with silkworm pupas removed; further, the silkworm cocoon is an integral open-end silkworm cocoon or a cut-up silkworm cocoon, wherein the cut-up silkworm cocoon is strip-shaped or square and has an area of 1-1000 mm2。
Preferably, the concentration of the sodium carbonate solution is 0-100 g/L, and the ratio of the mass of the silkworm cocoon to the volume of the sodium carbonate solution is 10: (1-100) g/L.
Preferably, the heating mode is electric furnace heating or pressure cooker heating, the heating temperature is 50-121 ℃, the heating pressure is 0.1-2 MPa, and the heating time is 1 s-1000 h.
Preferably, the average molecular weight of the silk fibroin is adjusted by controlling the heating time, and the average molecular weight of the silk fibroin is 100 KDa-350 KDa.
Preferably, the formed silk is washed in ultrapure water and repeated several times; further, the washing mode is stirring washing or standing soaking, wherein the stirring mode of the stirring washing is mechanical stirring or magnetic stirring, and the magnetic stirring speed is 1 r/min-2000 r/min; in a single washing process, the mass ratio of the silk to water is 1: 10-1: 1000, the time is 1 s-200 h, the silk is taken out and squeezed to be dry after each washing, and the washing repetition times are 1-50.
Preferably, the first drying is natural air drying or heating and ventilation drying, wherein the heating temperature of the heating and ventilation drying is 40-100 ℃, and the drying time is 0.1-100 h.
Preferably, the concentration of LiBr or LiSCN solution is 0.01-2 g/mL, and the volume ratio of the mass of the silk after primary drying to the LiBr or LiSCN solution is (1-100): 100 g/mL.
Preferably, the temperature for heat preservation is 20-100 ℃, and the preset time is 0.1-100 h.
Preferably, the dialysis bag for dialysis is 10-10000000 Da in specification, the dialysis mode is static dialysis or magnetic stirring dialysis, wherein the stirring speed of the magnetic stirring dialysis is 1-2000 r/min, the time interval of water change in the dialysis process is 0.1-100 h, and the volume of ultrapure water changed each time is 1 mL-1000L.
Preferably, the rotating speed of the centrifugation is 1-40000 r/min, the time is 1 s-10 h, and the temperature during the centrifugation is-3-10 ℃.
Preferably, the second freeze-drying comprises freezing the silk fibroin solution at a temperature of-80-0 ℃; freezing the frozen silk fibroin solution in vacuum; wherein the time is 1-500 hours, the vacuum pressure is 0.001-1 mBar, and the temperature is-80-0 ℃. Further, the vacuum freezing treatment is performed by using a freeze dryer.
In one embodiment of the invention, the average molecular weight of the silk fibroin is 100-350 KDa.
In one embodiment of the present invention, the step (1) comprises: dissolving 1 part by weight of the silk fibroin in 1-20 parts by weight of hexafluoroisopropanol; wherein the temperature of the silk fibroin and/or the hexafluoroisopropanol is 5-80 ℃, and the dissolving time is 0.1-100 hours.
In one embodiment of the present invention, the diameter of the opening is 0.05 to 2 mm, and the distribution density on the peripheral wall is 1 to 50/cm.
In one embodiment of the present invention, the protective shell is a plastic hard protective shell or a polymer film protective shell.
In an embodiment of the invention, the standing time is 1 to 100 hours.
In one embodiment of the present invention, the diameter of the water-soluble salt particles is 0.05 to 2 mm.
In one embodiment of the invention, the water-soluble salt particles are sodium chloride particles and/or potassium chloride particles.
In one embodiment of the present invention, the volume ratio of the water-soluble salt particles to the water-soluble salt particles-silk fibroin solution mixture is 1% to 99%.
In one embodiment of the present invention, the step (4) includes: the methanol is replaced every 1 to 50 hours, and the liquid replacement times are 1 to 50.
In one embodiment of the present invention, the step (4) includes: and removing the porous mold after the silk fibroin in the water-soluble salt particle-silk fibroin solution mixture is solidified.
In one embodiment of the present invention, the step (5) comprises: soaking the silk fibroin solid containing the water-soluble salt particles by using methanol aqueous solution with gradually decreased concentration gradient; wherein the concentration range of methanol in the methanol aqueous solution is 100-0, the replacement interval time range of the methanol aqueous solution is 1-50 hours, and the liquid replacement times are 1-50 times.
In an embodiment of the invention, the preparation method of the porous-solid composite silk material further comprises: drying the porous-solid composite silk fibroin material in a ventilated environment; wherein the temperature of the ventilation environment is 10-80 ℃, and the drying treatment time is 1-200 days.
In an embodiment of the invention, the preparation method of the porous-solid composite silk material further comprises: cutting the porous-solid composite silk fibroin material; wherein the cutting comprises turning and/or milling.
In one embodiment of the invention, the mechanical properties of the porous-solid composite silk material are regulated by controlling the average molecular weight of the silk fibroin.
The invention also provides a porous-solid composite silk material, which is prepared by the preparation method of the porous-solid composite silk material.
In a third aspect of the invention there is provided the use of the aforementioned porous-solid composite silk material in the manufacture of a bioimplantation medical device.
The fourth aspect of the invention provides the use of the aforementioned porous-solid composite silk material in a novel microelectronics field.
Compared with the prior art, the invention has the following beneficial effects:
the preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and is suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and pore diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical property, and can be widely applied to the field of bioimplantation medical treatment.
Drawings
Fig. 1 is a schematic flow chart of a preparation method of a porous-solid composite silk material provided by an embodiment of the invention.
Fig. 2 is a schematic diagram of a silk fibroin solution provided by an embodiment of the present invention.
Fig. 3 is a schematic diagram of a water-soluble salt particle-silk fibroin solution mixture provided by an embodiment of the present invention.
Fig. 4 is a schematic diagram of preparation of a silk fibroin solid containing water-soluble salt particles according to an embodiment of the present invention.
Fig. 5 is a schematic diagram of a silk fibroin solid containing water-soluble salt particles according to an embodiment of the present invention.
Fig. 6 is a schematic view of a porous-solid composite silk fibroin material provided by the embodiment of the present invention.
Fig. 7 is a schematic view of a porous-solid composite silk fibroin material formed by cutting according to an embodiment of the present invention.
Fig. 8 is a schematic view of a porous-solid composite silk fibroin material formed by cutting according to an embodiment of the present invention.
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 preparation method of a porous-solid composite silk material, and as shown in fig. 1, the preparation method of the porous-solid composite silk material comprises the following steps.
And S1, dissolving the silk fibroin in hexafluoroisopropanol to obtain the silk fibroin solution 1 shown in figure 2 (1 in figure 2 represents silk fibroin solution).
Specifically, 10g of silkworm cocoons are cut into pieces and placed in 4L of sodium carbonate solution with the concentration of 2g/L, the mixture is heated and boiled by an electric furnace and stirred by magnetic force, the rotating speed is 200r/min, and the heating time is 60 minutes. Then, each part of the boiled silk is put into 2L of ultrapure water to be stirred, washed and kneaded to be dry, the stirring is repeated for 6 times, and then the silk is dried, wherein the stirring speed is 200r/min, the stirring time is 20min, the volume of the ultrapure water changed each time is 2L, and the drying mode is that the silk is dried at 25 ℃ and normal pressure (0.1 MPa). Then, the dried silk is immersed in a lithium bromide solution with the concentration of 0.98g/mL, the volume of the lithium bromide solution is 4mL:1g relative to that of the dried silk, and the silk is kept at 60 ℃ for 4 hours to obtain a mixed solution of the fibroin and the lithium bromide. Then, the mixed solution of fibroin and lithium bromide is filled into a dialysis bag with molecular weight cutoff of 3500Da, and the dialysis bag is placed in 4L of ultrapure water for dialysis, and the ultrapure water is replaced once at intervals of 0.5h, wherein the volume of each water replacement is 4L, and the total dialysis time is 48 h. And finally, centrifuging the dialyzed fibroin solution at 4 ℃, rotating at 12000r/min for 30min, and collecting supernatant to finally obtain the required fibroin aqueous solution.
Then, the aqueous fibroin solution obtained above was dried. Specifically, freezing was performed using a-80 ℃ refrigerator, or freezing was performed using liquid nitrogen. The freeze-vacuum condition is that a vacuum drier is used, and the vacuum freeze treatment is carried out in the freeze drier for 100 hours, wherein the vacuum pressure of the freeze drier is 0.02mBar, and the temperature is-10 ℃. Obtaining powdery silk fibroin.
The mass ratio of the powdery silk fibroin to the hexafluoroisopropanol is 1: 4, the dissolution is carried out at a dissolution environment temperature range of 25 ℃ (room temperature environment) and a dissolution time range of 24 hours. When dissolving, it is necessary to seal the container to prevent volatilization of hexafluoroisopropanol, and a sealed glass bottle with a cap, a sealed syringe, a beaker sealed with a polymer film, and the like are used.
S2, injecting the silk fibroin solution into a porous mold 2 shown in figure 3 (2 in figure 3 represents a porous mold), and standing; wherein the porous mold 2 includes a peripheral wall having a plurality of openings and a protective shell 3 (3 in fig. 3 represents a protective shell) as shown in fig. 3.
Specifically, silk fibroin solution 1 is injected into a porous mold 2. The porous mold 2 comprises a protective shell 3, which prevents the silk fibroin solution 1 from flowing out. The protective shell may be a plastic hard protective shell, or a polymer film protective shell, but is not limited thereto. The peripheral wall of the porous mold 2 is provided with a plurality of openings, the diameter range of the openings is 0.05-2 mm, and the density range of the openings is 1-50 per square centimeter. And (3) after the silk fibroin solution 1 is injected into the porous mold 2, standing for 24 hours until the bubbles in the solution are completely released.
S3, filling the silk fibroin solution in the porous mold 2 with water-soluble salt particles 4 (4 in fig. 3 represents water-soluble salt particles) as shown in fig. 3 to obtain a water-soluble salt particle-silk fibroin solution mixture; wherein the water-soluble salt particles are insoluble in hexafluoroisopropanol.
The water-soluble salt particles 4 are allowed to stand for a while to be sufficiently precipitated, and the filling volume ratio of the water-soluble salt particles 4 is controlled. Specifically, sodium chloride particles 4 (average diameter of 0.5 mm) were filled in a hexafluoroisopropanol solution of silk fibroin, left for 12 hours to sufficiently precipitate sodium chloride, and the volume ratio of sodium chloride in the solution was controlled to 50%.
S4, soaking the porous mold 3 containing the water-soluble salt particle-silk fibroin solution mixture after removing the protective shell 3 in methanol 5 (5 in fig. 4 represents methanol) shown in fig. 4 to obtain silk fibroin solid 6 containing water-soluble salt particles shown in fig. 5 (6 in fig. 5 represents silk fibroin solid with water-soluble salt particles).
At S4, the hexafluoroisopropanol is replaced with methanol 5 while the silk fibroin crystals are solidified, and the silk fibroin crystals can be air-dried in a ventilated environment to obtain silk fibroin solids 6 containing water-soluble salt particles 4.
Specifically, after the air bubbles of the silk fibroin solution 1 are completely released, the protective shell 3 covered on the surface of the porous mold 2 is removed, and the porous mold 2 containing the silk fibroin solution 1 and the sodium chloride salt particles 4 is soaked in a methanol 5 environment. The methanol solution was changed every 12 hours for 10 times. And (3) completely treating and solidifying the silk fibroin by methanol, removing the porous mold 2, volatilizing moisture in the silk fibroin in a ventilation environment, and airing, wherein the ventilation environment temperature is 25 ℃, and the treatment time is 20 days to obtain the silk fibroin solid 6 containing the water-soluble salt particles 4.
S5, removing the methanol and the water-soluble salt particles 4 in the silk fibroin solid 6 containing the water-soluble salt particles to obtain the porous-solid composite silk fibroin material.
The methanol and water-soluble salt particles 4 are removed by displacement. Specifically, the silk fibroin solid 6 containing the water-soluble salt particles 4 is soaked in an aqueous methanol solution 7 (7 in fig. 6 represents an aqueous methanol solution) as shown in fig. 6, in which the concentration gradient decreases; wherein, the concentration of the methanol in the methanol water solution 7 is reduced from 100 percent to 0 percent in turn, and the methanol water solution 7 is replaced once every 12 hours; the number of liquid changes was 10. The remaining methanol is replaced with water by the replacement method, and the water-soluble salt particles 4 are sufficiently dissolved in water.
And (3) drying the porous-solid composite silk fibroin material obtained in the S5 in a ventilated environment, and cutting and modifying the dried porous-solid composite silk fibroin material into a required shape, wherein the shape can be a shape represented by 8, 9 and 10 in the figure 7.
In one example, porous-solid composite silk fibroin materials are dried in a ventilated environment, specifically, the moisture in the silk fibroin solids is volatilized and air dried in a ventilated environment, the temperature of the ventilated environment is 25 ℃, and the processing time is 15 days. The cutting and modifying process includes turning, milling and other methods, but is not limited to the method; the shape of the cut porous-solid composite silk fibroin material comprises a cuboid, a cube, a cylinder and a triangle, but is not limited to the shape; the size range of the shape of the cut porous-solid composite silk fibroin material is 0.1-1000 cubic centimeters. Figure 8 shows a porous-solid composite silk fibroin cylinder material that is cut.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and is suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and pore diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical property, and can be widely applied to the field of bioimplantation medical treatment.
The embodiment of the invention also provides the porous-solid composite silk material prepared by the preparation method of the porous-solid composite silk material.
The porous-solid composite silk material provided by the embodiment of the invention has adjustable porosity and pore diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties, and can be widely applied to the field of bioimplantation medical treatment.
Example 2
In example 2, the lithium bromide solution in S1 was replaced with a lithium thiocyanate solution, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 3
In example 3, the mass ratio of powdered silk fibroin and hexafluoroisopropanol was 1:1, the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 4
In this example, the mass ratio of powdered silk fibroin to hexafluoroisopropanol was 1: the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 5
In this example, the mass ratio of powdered silk fibroin to hexafluoroisopropanol was 1: 20, the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 6
In this example, powdered silk fibroin was dissolved in hexafluoroisopropanol at a temperature of 5 ℃ for 50 hours, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 7
In this example, powdered silk fibroin was dissolved in hexafluoroisopropanol at 40 ℃ for 0.1 hour, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 8
In this example, powdered silk fibroin was dissolved in hexafluoroisopropanol at 80 ℃ for 100 hours, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 9
In this example, the silk fibroin solution was injected into a porous mold and then allowed to stand for 100 hours, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 10
In this example, the silk fibroin solution was injected into a porous mold and then allowed to stand for 50 hours, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 11
In this example, the silk fibroin solution was injected into a porous mold and then left to stand for 1 hour, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 12
In this example, the water-soluble salt particles were potassium chloride, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 13
In this example, the water-soluble salt particles were potassium chloride, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 14
In this example, the volume ratio of sodium chloride in the solution was controlled to 99%, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 15
In this example, the volume ratio of sodium chloride in the solution was controlled to 1%, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 16
In this example, methanol was preferably added every 1 hour in S4, and the number of liquid changes was 25, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 17
In this example, methanol was preferably added every 25 hours in S4, the number of liquid changes was 1, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 18
In this example S4, methanol was replaced every 50 hours, the number of times of liquid replacement was 25, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 19
In this example S5, methanol was replaced every 50 hours, the number of times of liquid replacement was 25, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 20
In this example S4, methanol was preferably added every 25 hours, the number of liquid changes was 50, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 21
In this example S4, methanol was preferably added every 1 hour, and the number of liquid changes was 10, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 22
The porous-solid composite silk fibroin material obtained in S5 was dried in a ventilated environment at a temperature of 80 ℃ for 100 days, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 23
The porous-solid composite silk fibroin material obtained in S5 was dried for 50 days in a ventilated environment at a temperature of 40 ℃, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
Example 24
The porous-solid composite silk fibroin material obtained in S5 was dried in a ventilated environment at a temperature of 10 ℃ for 200 days, and the other operations were the same as in example 1.
The preparation method of the porous-solid composite silk material provided by the embodiment of the invention is simple in process operation and suitable for industrialization; the prepared porous-solid composite silk material has adjustable porosity and gap diameter, firm combination of the porous structure and the solid structure, and adjustable biodegradation and mechanical properties.
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 (18)
1. The preparation method of the porous-solid composite silk material is characterized by comprising the following steps of:
(1) dissolving silk fibroin in hexafluoroisopropanol to obtain a silk fibroin solution;
(2) injecting the silk fibroin solution into a porous mold, and standing; wherein the porous mold comprises a peripheral wall and a protective shell, the peripheral wall having a plurality of openings;
(3) filling water-soluble salt particles into the silk fibroin solution in the porous mold to obtain a water-soluble salt particle-silk fibroin solution mixture;
(4) soaking the porous mold containing the water-soluble salt particle-silk fibroin solution mixture after the protective shell is removed in methanol to prepare silk fibroin solid containing water-soluble salt particles;
(5) and removing methanol and water-soluble salt particles in the silk fibroin solid containing the water-soluble salt particles to obtain the porous-solid composite silk fibroin material.
2. The method for preparing the porous-solid composite silk material of claim 1, wherein in the step (1), the silk fibroin is anhydrous silk fibroin.
3. The method for preparing porous-solid composite silk material according to claim 1, wherein the silk fibroin is anhydrous silk fibroin dry powder obtained by direct freeze drying method from aqueous solution state before dissolving in hexafluoroisopropanol;
and dissolving the anhydrous silk fibroin dry powder in hexafluoroisopropanol to obtain the silk fibroin solution.
4. The method for preparing a porous-solid composite silk material according to claim 1, wherein the step (1) comprises:
placing the silkworm cocoon in a sodium carbonate solution, and heating to form silk; first drying; adding LiBr or LiSCN solution, and preserving heat for preset time to obtain mixed solution;
dialyzing the mixed solution, centrifuging the dialyzed solution, and taking supernatant to obtain a silk fibroin aqueous solution; and secondly, freeze-drying the silk fibroin aqueous solution to obtain the silk fibroin.
5. The method for preparing a porous-solid composite silk material according to claim 4, wherein the second freeze-drying is:
freezing the silk fibroin aqueous solution at the temperature of-80-0 ℃;
freezing the frozen silk fibroin aqueous solution in vacuum; wherein the time is 1-500 hours, the vacuum pressure is 0.001-1 mBar, and the temperature is-80-0 ℃.
6. The method for preparing the porous-solid composite silk material according to claim 1, wherein the average molecular weight of the silk fibroin is 100-350 KDa.
7. The method for preparing a porous-solid composite silk material according to claim 1, wherein the step (1) comprises:
dissolving 1 part by weight of the silk fibroin in 1-20 parts by weight of hexafluoroisopropanol; wherein the temperature of the silk fibroin and/or the hexafluoroisopropanol is 5-80 ℃, and the dissolving time is 0.1-100 hours.
8. The method for preparing a porous-solid composite silk material according to claim 1, wherein the diameter of the open pores is 0.05-2 mm, and the distribution density on the peripheral wall is 1-50 per square centimeter.
9. The method for preparing a porous-solid composite silk material according to claim 1, wherein the protective shell is a plastic hard protective shell or a polymer film protective shell.
10. The method for preparing the porous-solid composite silk material according to claim 1, wherein the standing time is 1-100 hours.
11. The method for preparing a porous-solid composite silk material according to claim 1, wherein the water-soluble salt particles have a diameter of 0.05-2 mm.
12. The method for preparing a porous-solid composite silk material according to claim 1, wherein the water-soluble salt particles are sodium chloride particles and/or potassium chloride particles.
13. The method for preparing porous-solid composite silk material according to claim 1, wherein the volume ratio of the water-soluble salt particles to the water-soluble salt particle-silk fibroin solution mixture is 1-99%.
14. The method for preparing a porous-solid composite silk material according to claim 1, wherein the step (4) comprises:
the methanol is replaced every 1 to 50 hours, and the liquid replacement times are 1 to 50.
15. The method for preparing a porous-solid composite silk material according to claim 1, wherein the step (4) comprises: and removing the porous mold after the silk fibroin in the water-soluble salt particle-silk fibroin solution mixture is solidified.
16. The method for preparing a porous-solid composite silk material according to claim 1, wherein the step (5) comprises:
soaking the silk fibroin solid containing the water-soluble salt particles by using methanol aqueous solution with gradually decreased concentration gradient; wherein the concentration range of methanol in the methanol aqueous solution is 100-0, the replacement interval time range of the methanol aqueous solution is 1-50 hours, and the liquid replacement times are 1-50 times.
17. The method for preparing porous-solid composite silk material according to claim 1, further comprising:
drying the porous-solid composite silk fibroin material in a ventilated environment; wherein the temperature of the ventilation environment is 10-80 ℃, and the drying treatment time is 1-200 days.
18. The method for preparing porous-solid composite silk material according to claim 1, further comprising:
cutting the porous-solid composite silk fibroin material; wherein the cutting comprises turning and/or milling.
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| CN101406713A (en) * | 2007-10-12 | 2009-04-15 | 微创医疗器械(上海)有限公司 | Artificial blood vessel bracket and preparation method thereof |
| CN101596327A (en) * | 2009-07-13 | 2009-12-09 | 北京理工大学 | A kind of preparation method of three-dimensional silk fibroin porous scaffold material |
| CN101857729A (en) * | 2010-06-02 | 2010-10-13 | 苏州大学 | Water-insoluble silk fibroin porous material and preparation method thereof |
| CN102430155A (en) * | 2011-12-08 | 2012-05-02 | 西安交通大学 | Cellular silk fibroin porous scaffold, and preparation method thereof |
| CN102512710A (en) * | 2011-12-28 | 2012-06-27 | 苏州大学 | Preparation method of porous three-dimensional silk fibroin material |
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| CN101406713A (en) * | 2007-10-12 | 2009-04-15 | 微创医疗器械(上海)有限公司 | Artificial blood vessel bracket and preparation method thereof |
| CN101596327A (en) * | 2009-07-13 | 2009-12-09 | 北京理工大学 | A kind of preparation method of three-dimensional silk fibroin porous scaffold material |
| CN101857729A (en) * | 2010-06-02 | 2010-10-13 | 苏州大学 | Water-insoluble silk fibroin porous material and preparation method thereof |
| CN102430155A (en) * | 2011-12-08 | 2012-05-02 | 西安交通大学 | Cellular silk fibroin porous scaffold, and preparation method thereof |
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