CN111041603A - Preparation method of fibroin/microorganism-based polymer solution and preparation method of composite nanofiber of fibroin/microorganism-based polymer solution - Google Patents
Preparation method of fibroin/microorganism-based polymer solution and preparation method of composite nanofiber of fibroin/microorganism-based polymer solution Download PDFInfo
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- CN111041603A CN111041603A CN201911333145.2A CN201911333145A CN111041603A CN 111041603 A CN111041603 A CN 111041603A CN 201911333145 A CN201911333145 A CN 201911333145A CN 111041603 A CN111041603 A CN 111041603A
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
- D01F8/00—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof
- D01F8/02—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof from cellulose, cellulose derivatives, or proteins
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D1/00—Treatment of filament-forming or like material
- D01D1/02—Preparation of spinning solutions
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01F—CHEMICAL FEATURES IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS; APPARATUS SPECIALLY ADAPTED FOR THE MANUFACTURE OF CARBON FILAMENTS
- D01F8/00—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof
- D01F8/04—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof from synthetic polymers
- D01F8/14—Conjugated, i.e. bi- or multicomponent, artificial filaments or the like; Manufacture thereof from synthetic polymers with at least one polyester as constituent
Abstract
The invention discloses a preparation method of silk protein/microorganism-based polymer solution and a preparation method of composite nano-fiber thereof, wherein formic acid and trichloromethane are selected as solvents, 98 percent of purity formic acid is used for dissolving silk protein, and trichloromethane is used for dissolving PHBV; mixing the silk protein solution and the PHBV solution according to a certain solute mass ratio by regulating the mass fractions of the silk protein solution and the PHBV solution to obtain a stable spinnable mixed spinning solution; finally, the fibroin/PHBV composite nano-fiber is prepared by a needle type electrostatic spinning method. The fibroin/PHBV composite nanofiber prepared by the method is simple in preparation method, the fibroin/PHBV composite nanofiber simultaneously contains two components of fibroin and PHBV, and the diameter of the fiber is 92.44 nm-1273.15 nm.
Description
Technical Field
The invention belongs to the technical field of nano-fibers, and particularly relates to a preparation method of a fibroin/microorganism-based polymer solution and a preparation method of composite nano-fibers thereof.
Background
Over the past few decades, nanomaterials have attracted considerable attention as nanotechnology has emerged and continues to advance. The electrostatic spinning technology is a simple and effective common method for preparing superfine nano fibers by electric field force, and the fiber diameter is generally between dozens of nanometers and several micrometers. In the electrostatic spinning process, the jet flow at the spinning needle moves rapidly towards the receiving plate under the action of the electric field force, during which the solvent in the spinning solution will volatilize and eventually form fibers on the receiving plate. Compared with the traditional fiber, the electrostatic spinning nanofiber has the characteristics of superfine property, large specific surface area, high porosity and the like, and can be widely applied to the fields of tissue engineering, protective clothing, filtration, electronic devices and the like. It is worth noting that in the future biomedical field, the electrostatic spinning nano material will play a significant role.
Silkworm-raising and silk-reeling have a long history in China, and because silk has special luster and outstanding physical and mechanical properties, people particularly like clothes made of silk from ancient times to present. In the past, silk has been used primarily in the apparel industry. In recent years, the production of biomedical materials from silk has been increasingly studied.
Studies on silk protein complex fibers, such as: firstly, preparing a regenerated silk fibroin film, then synthesizing silver nanowires, dissolving the prepared silk fibroin film in a silver nanowire/acid solvent dispersion system to form silk fibroin/silver nanowires/acid solvent spinning solution, and finally preparing single-fiber silver nanowire/silk fibroin composite fibers with high conductivity and high flexibility; for another example: firstly, obtaining silk fibroin and hydroxypropyl methyl cellulose, dissolving the silk fibroin and the hydroxypropyl methyl cellulose in formic acid to obtain a basic spinning solution, placing the basic spinning solution in a magnetic stirrer to be stirred to be a uniform and transparent solution, then placing the spinning solution in a constant liquid supply device, controlling the liquid supply speed of the constant liquid supply device to form a free liquid level on the upper surface of a liquid storage pipe, and opening a high-voltage electrostatic device to start spinning to obtain the nano-fibers.
In addition, with the continuous development of scientific technology, people pay more attention to environmental protection, and the use of environment-friendly materials is promoted more and more. PHA is intracellular polyester synthesized by microorganisms such as bacteria, and is a reserve material of carbon sources and energy sources in bacterial cells. Silk protein (SF) and Polyhydroxyalkanoates (PHA) have unique biocompatibility and biodegradability and have received increasing attention in recent years. The microbial-based polymer material used in the present invention is poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), which is one of many kinds of PHA, and is copolymerized from 3-hydroxybutyrate (PHB) and 3-Hydroxyvalerate (HV). At present, the existing preparation method of the spinning solution for preparing the fibroin/PHBV composite nanofiber adopts hexafluoroisopropanol as a common solvent of silk fibroin and PHBV, but hexafluoroisopropanol has acute toxicity and high price, and has great potential hazard in the use process.
Therefore, it is necessary to provide a further solution to the above existing problems.
Disclosure of Invention
The invention aims to provide a preparation method of a silk protein/microorganism-based polymer solution and a preparation method of composite nano-fibers thereof, and solves the problems.
The technical scheme of the invention is as follows:
a method of preparing a fibroin/microbe-based polymer solution, the method comprising the steps of:
(1) preparation of silk protein solution: cutting a silk fibroin film into pieces, dissolving the silk fibroin film by formic acid to obtain a silk fibroin solution, and placing the silk fibroin solution in a refrigerator for later use;
(2) preparing a PHBV solution: dissolving PHBV by using trichloromethane, and stirring until the PHBV is completely dissolved to obtain a PHBV solution;
(3) preparing a spinning solution: and mixing the silk protein solution and the PHBV solution to obtain spinning solution, namely silk protein/microorganism-based polymer solution.
Further, the preparation method of the silk fibroin membrane in the step (1) comprises the following steps: boiling raw silk with 0.02mol/L of Na2CO3Degumming in the solution for 30min to obtain the degummed silk fibroin; putting the degummed silk fibroin in an oven at 60 DEG CDrying, and then dissolving the dried silk fibroin into 9.3mol/L LiBr solution to obtain a silk fibroin solution; dialyzing the fibroin solution for 3 days, centrifuging by using a centrifugal machine, repeating for 2 times, wherein the parameters are 12000rpm, 4 ℃ and 20min, and filtering to obtain a fibroin aqueous solution; and dripping the fibroin aqueous solution into a culture dish, and drying in a 35 ℃ drying oven to finally obtain the fibroin membrane.
Further, the addition ratio of the dried silk fibroin to the LiBr solution is as follows: 13.5g of dried silk fibroin was dissolved in 50ml of LiBr solution.
Further, the purity of the formic acid in the step (1) is 98%.
Further, the mass percent of the fibroin solution in the step (1) is 8% -10%.
Further, the stirring temperature in the step (2) is constant at 50 ℃.
Further, the PHBV solution in the step (2) is 2-5% by mass.
Further, the solute mass ratio of the fibroin and the PHBV of the spinning solution in the step (3) is any one of 1.5/1, 2/1, 3/1, 4/1, 5/1, 6/1, 7/1, 8/1, 9/1 and 10/1.
The other technical scheme of the invention is as follows:
a preparation method of composite nanofibers utilizes the silk protein/microorganism-based polymer solution prepared by the method to prepare the composite nanofibers by an electrostatic spinning method.
Further, a needle type electrostatic spinning device is adopted in the electrostatic spinning method, a metal flat plate is used as a collecting device, the spinning temperature is room temperature, the humidity is less than 40%, the spinning voltage is 15-23 kV, the flow is 0.3mL/h, and the receiving distance is 13-17 cm.
The invention provides a preparation method of a silk protein/microorganism-based polymer solution and a preparation method of composite nano-fibers thereof, and the method has the following advantages:
1. compared with pure silk protein fibers and pure PHBV fibers, the prepared silk protein/microorganism-based polymer composite nanofibers can regulate and control the fiber diameter between dozens of nanometers and micrometers according to different mixed spinning solutions, thereby meeting the requirements of different applications;
2. the method has simple process and easy operation;
3. the silk protein/microorganism-based polymer composite nanofiber prepared by the method has good biodegradability and biocompatibility, in addition, the PHBV can improve the crystallinity of the composite nanofiber, and the silk protein can improve the thermal property of the composite nanofiber, so that the silk protein/microorganism-based polymer composite nanofiber has the common advantages of the two materials.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the description of the embodiments will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings based on these drawings without inventive exercise. Wherein the content of the first and second substances,
FIG. 1 is an electron microscope image of appearance morphology of silk fibroin/microbial-based polymer composite nanofibers prepared with different PHBV contents according to the present invention, wherein (a) the solute mass ratio of silk fibroin to PHBV in the image is 10/1, (b) the solute mass ratio of silk fibroin to PHBV in the image is 4/1, (c) the solute mass ratio of silk fibroin to PHBV in the image is 3/1, (d) the solute mass ratio of silk fibroin to PHBV in the image is 2/1, (e) the solute mass ratio of silk fibroin to PHBV in the image is 1.5/1, and (f) the solute mass ratio of silk fibroin to PHBV in the image is 1.5/1, but the fiber morphology is significantly different from that of (e).
Detailed Description
The invention discloses a preparation method of a silk protein/microorganism-based polymer solution, which uses two solvents, wherein the two solvents are common chemical reagents and are low in price. Dissolving silk protein with formic acid, dissolving PHBV with chloroform, mixing the silk protein solution and PHBV solution according to a certain solute mass ratio, and finally obtaining uniform and stable mixed spinning solution. The specific operation steps are as follows:
(1) preparation of silk protein solution: the silk fibroin solution is obtained by dissolving a silk fibroin membrane with formic acid. The silk fibroin film is cut into pieces, 98 percent of purity formic acid is adopted to dissolve the silk fibroin film, the mass fraction of the silk fibroin solution is 8 percent to 10 percent, and the silk fibroin solution is put in a refrigerator at 4 ℃ for standby.
(2) Preparation of PHBV solution: dissolving PHBV with trichloromethane, and stirring at a constant temperature of 50 ℃ until the PHBV is completely dissolved, wherein the mass fraction of the PHBV solution is 2-5%.
(3) The preparation of spinning solution comprises the steps of mixing the two solutions obtained according to different solute mass ratios of silk fibroin and PHBV in the mixed solution, wherein the mixing ratio of the two solutions is 1/1, 2/1, 3/1, 4/1, 5/1, 6/1, 7/1, 8/1, 9/1 and 10/1, and the mixed spinning solution is found to generate precipitates in the process of preparing the mixed spinning solution mainly due to the facts that trichloromethane serving as an organic solvent can reduce the dielectric constant of a protein solution and accordingly increase electrostatic repulsive force in protein molecules and reduce stability, trichloromethane ② belongs to halogenated hydrocarbons and can react with amino groups in proteins to form amines and cause protein denaturation and precipitation, the concentration of the silk protein solution is reduced, the concentration of the PHBV solution is increased, the content of PHBV in the stable spinnable mixed spinning solution can be effectively increased, therefore, when the mixed spinning solution is prepared, the mixing ratio of silk fibroin and PHBV needs to be well controlled, and the mixed spinning solution with the solutes of silk fibroin and PHBV is adjusted, the mixed spinning solution is prepared, and the mixed spinning solution is capable of obtaining stable spinning solution with stable spinning solution quality ratios of 354642, 3527, 3684 and stable fibers, and 5/1, and 3684 and 3642 are obtained.
The spinning solution is utilized, a needle type electrostatic spinning device and a metal flat plate are used as a collecting device, the spinning temperature is room temperature, the humidity is below 40%, the spinning voltage is 15-23 kV, the flow rate is 0.3mL/h, the receiving distance is 13-17 cm, and the composite nanofiber is prepared. The spinning parameters have great influence on the spinning state, the fiber morphology and the like of the composite nanofiber.
In order to make the aforementioned objects, features and advantages of the present invention more comprehensible, embodiments accompanying the present invention are further described below. The invention is not limited to the embodiments listed but also comprises any other known variations within the scope of the invention as claimed.
Reference herein to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic may be included in at least one implementation of the invention. The appearances of the phrase "in one embodiment" in various places in the specification are not necessarily all referring to the same embodiment, nor are separate or alternative embodiments mutually exclusive of other embodiments.
Example 1
The embodiment shows a preparation method of the composite nanofiber according to the following steps, and the specific steps are as follows:
(1) preparing a silk fibroin film: boiling raw silk with 0.02mol/L of Na2CO3Degumming the solution for 30min, drying the degummed silk fibroin in an oven at 60 ℃, dissolving the dried silk fibroin in 9.3mol/L LiBr solution (the addition ratio: 13.5g silk fibroin is dissolved in 50ml LiBr solution), dialyzing the obtained silk fibroin solution for 3 days, centrifuging by using a centrifuge, repeating the steps for 2 times, wherein the parameters are 12000rpm, the temperature is 4 ℃, and the time is 20min, and filtering to obtain the silk fibroin aqueous solution. And dripping the silk fibroin aqueous solution into a culture dish, and drying in a 35 ℃ drying oven to finally obtain the silk fibroin membrane.
(2) Preparing a spinning solution: dissolving the fibroin membrane obtained in the step (1) by using 98% purity formic acid to obtain a fibroin solution with the mass fraction of 10%; dissolving PHBV with chloroform to obtain a PHBV solution with the mass fraction of 2%, mixing the two solutions according to the mass ratio of the fibroin to the PHBV as 10/1, and spinning with the mixed spinning solution.
(3) Preparing the composite nanofiber: the needle type electrostatic spinning device is adopted in the experiment, a metal flat plate is used as a receiving device, the air humidity is below 40%, the spinning specific parameters are voltage 15kV, flow rate 0.3mL/h, receiving distance 13cm, and fiber diameter 171.10 +/-40.38 nm.
Example 2
The embodiment shows a preparation method of the composite nanofiber according to the following steps, and the specific steps are as follows:
(1) preparing a silk fibroin film: the procedure was the same as in step (1) of example 1.
(2) Preparing a spinning solution: dissolving the fibroin membrane obtained in the step (1) by using 98% purity formic acid to obtain a fibroin solution with the mass fraction of 10%; dissolving PHBV with chloroform to obtain a PHBV solution with the mass fraction of 3%, mixing the two solutions according to the mass ratio of the fibroin to the PHBV solute of 4/1, and spinning.
(3) Preparing the composite nanofiber: the needle type electrostatic spinning device is adopted in the experiment, a metal flat plate is used as a receiving device, the air humidity is below 40%, and the spinning parameters are 20kV voltage, 0.3mL/h flow, 17cm receiving distance and 149.70 +/-28.57 nm fiber diameter.
Example 3
The embodiment shows a preparation method of the composite nanofiber according to the following steps, and the specific steps are as follows:
(1) preparing a silk fibroin film: the procedure was the same as in step (1) of example 1.
(2) Preparing a spinning solution: dissolving the fibroin membrane obtained in the step (1) by using 98% purity formic acid to obtain a fibroin solution with the mass fraction of 8%; dissolving PHBV by using trichloromethane to obtain a PHBV solution with the mass fraction of 4%, and preparing a mixed solution for spinning when the mass ratio of the fibroin to the PHBV is 3/1.
(3) Preparing the composite nanofiber: the needle type electrostatic spinning device is adopted in the experiment, a metal flat plate is used as a receiving device, the air humidity is below 40%, and the spinning specific parameters are voltage 23kV, flow rate 0.3mL/h, receiving distance 13cm and fiber diameter 108.29 +/-26.77 nm.
Example 4
The embodiment shows a preparation method of the composite nanofiber according to the following steps, and the specific steps are as follows:
(1) preparing a silk fibroin film: the procedure was the same as in step (1) of example 1.
(2) Preparing a spinning solution: dissolving the fibroin membrane obtained in the step (1) by using 98% purity formic acid to obtain a fibroin solution with the mass fraction of 8%; dissolving PHBV with chloroform to obtain a PHBV solution with the mass fraction of 4.5%, and preparing a mixed solution of the two solutions according to the mass ratio of the fibroin to the PHBV as 2/1 for spinning.
(3) Preparing the composite nanofiber: the needle type electrostatic spinning device is adopted in the experiment, a metal flat plate is used as a receiving device, the air humidity is below 40%, and the spinning parameters are 20kV voltage, 0.3mL/h flow, 17cm receiving distance and 120.82 +/-24.83 nm fiber diameter.
Example 5
The embodiment shows a preparation method of the composite nanofiber according to the following steps, and the specific steps are as follows:
(1) preparing a silk fibroin film: the procedure was the same as in step (1) of example 1.
(2) Preparing a spinning solution: dissolving the fibroin membrane obtained in the step (1) by using 98% purity formic acid to obtain a fibroin solution with the mass fraction of 8%; dissolving PHBV with chloroform to obtain a PHBV solution with the mass fraction of 5%, and preparing a mixed solution of the two solutions according to the mass ratio of fibroin to the solute of PHBV of 1.5/1 for spinning.
(3) Preparing the composite nanofiber: the needle type electrostatic spinning device is adopted in the experiment, a metal flat plate is used as a receiving device, the air humidity is below 40%, and the specific spinning parameters are voltage 20kV, flow rate 0.3mL/h, receiving distance 13cm and fiber diameter 92.44 +/-18.63 nm.
Example 6
The embodiment shows a preparation method of the composite nanofiber according to the following steps, and the specific steps are as follows:
(1) preparing a silk fibroin film: the procedure was the same as in step (1) of example 1.
(2) Preparing a spinning solution: dissolving the fibroin membrane obtained in the step (1) by using 98% purity formic acid to obtain a fibroin solution with the mass fraction of 8%; dissolving PHBV with chloroform to obtain a PHBV solution with the mass fraction of 4.5%, and preparing a mixed solution of the two solutions according to the mass ratio of fibroin to the solute of PHBV of 1.5/1 for spinning.
(3) Preparing the composite nanofiber: the needle type electrostatic spinning device is adopted in the experiment, a metal flat plate is used as a receiving device, the air humidity is below 40%, and the spinning parameters are 20kV voltage, 0.3mL/h flow, 13cm receiving distance and 1273.15 +/-225.18 nm fiber diameter.
Compared with the prior art, the invention has the beneficial effects that: the invention relates to a preparation method of silk protein/microorganism-based polymer solution and a preparation method of composite nanofiber thereof, which adopt a simple needle type electrostatic spinning technology and select two biological materials of silk protein and PHBV as raw materials to prepare the composite nanofiber with good biocompatibility and biodegradability. In addition, in the electrostatic spinning process, the crystallization capacity of silk protein is low, the silk protein is in a random coil structure, and PHBV is easy to crystallize, so that the crystallinity of the composite nanofiber can be effectively improved.
It should be noted that the above-mentioned embodiments are only for illustrating the technical solutions of the present invention and not for limiting, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions may be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention, which should be covered by the claims of the present invention.
Claims (10)
1. A method of preparing a fibroin/microbe-based polymer solution, comprising the steps of:
(1) preparation of silk protein solution: cutting a silk fibroin film into pieces, dissolving the silk fibroin film by formic acid to obtain a silk fibroin solution, and placing the silk fibroin solution in a refrigerator for later use;
(2) preparing a PHBV solution: dissolving PHBV by using trichloromethane, and stirring until the PHBV is completely dissolved to obtain a PHBV solution;
(3) preparing a spinning solution: and mixing the silk protein solution and the PHBV solution to obtain spinning solution, namely silk protein/microorganism-based polymer solution.
2. A silk protein/microbe-based polymer solution as defined in claim 1The preparation method is characterized by comprising the following steps: the preparation method of the silk fibroin film in the step (1) comprises the following steps: boiling raw silk with 0.02mol/L of Na2CO3Degumming in the solution for 30min to obtain the degummed silk fibroin; drying the degummed silk fibroin in a drying oven at 60 ℃, and then dissolving the dried silk fibroin in 9.3mol/L LiBr solution to obtain a silk fibroin solution; dialyzing the fibroin solution for 3 days, centrifuging by using a centrifugal machine, repeating for 2 times, wherein the parameters are 12000rpm, 4 ℃ and 20min, and filtering to obtain a fibroin aqueous solution; and dripping the fibroin aqueous solution into a culture dish, and drying in a 35 ℃ drying oven to finally obtain the fibroin membrane.
3. The method of claim 2, wherein the step of preparing the silk protein/microorganism-based polymer solution comprises: the addition ratio of the dried silk fibroin to the LiBr solution is as follows: 13.5g dried silk fibroin was dissolved in 50ml LiBr solution.
4. The method of claim 1, wherein the step of preparing the silk protein/microbe-based polymer solution comprises: the purity of the formic acid in the step (1) is 98%.
5. The method of claim 1, wherein the step of preparing the silk protein/microbe-based polymer solution comprises: the mass percentage of the fibroin solution in the step (1) is 8-10%.
6. The method of claim 1, wherein the step of preparing the silk protein/microbe-based polymer solution comprises: the stirring temperature in the step (2) is constant at 50 ℃.
7. The method of claim 1, wherein the step of preparing the silk protein/microbe-based polymer solution comprises: the mass percentage of the PHBV solution in the step (2) is 2-5%.
8. The method of claim 1, wherein the step of preparing the silk protein/microbe-based polymer solution comprises: the mass ratio of the solute of the fibroin and the PHBV in the spinning solution in the step (3) is any one of 1.5/1, 2/1, 3/1, 4/1, 5/1, 6/1, 7/1, 8/1, 9/1 and 10/1.
9. A method for preparing composite nano-fiber is characterized in that: preparing composite nanofibers by electrospinning using the fibroin/microbe-based polymer solution prepared according to any one of claims 1 to 8.
10. The method of claim 9, wherein the step of preparing the composite nanofiber comprises: the electrostatic spinning method adopts a needle type electrostatic spinning device, a metal flat plate is used as a collecting device, the spinning temperature is room temperature, the humidity is less than 40%, the spinning voltage is 15-23 kV, the flow is 0.3mL/h, and the receiving distance is 13-17 cm.
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Application publication date: 20200421 |