CN104250861B - A kind of electrospinning process prepared containing azelon - Google Patents

Abstract

The present invention relates to a kind of electrospinning process prepared containing azelon, protein-contg polymer solution is carried out electrospinning by described method in the atmosphere of crosslinking agent and water, be cross-linked while Electrospun is formed to realize protein compound, such cross-linking method can avoid cross-linking reaction on the impact of Electrospun pattern well, to solve in prior art crosslinked after fused mutually and reduce the problem of material specific surface area between Electrospun.Method is simple to operation, applied widely.Can be used for the electrostatic spinning containing protein polymer solution of protein solution, protein-charged non-protein polymer mixed liquor and the functional material (as nano material) that adulterates.

Description

An electrospinning method for preparing protein-containing fibers

technical field

The invention relates to an electrospinning method, in particular to an electrospinning method for making protein-containing fibers.

Background technique

Electrospinning is a method of preparing nano or micro polymer fibers. The fiber membrane obtained by the method has large specific surface area and high porosity, and has good application prospects in many fields. There are many polymers that can be electrospun, and the electrospinning of protein-like natural polymers is affected by tissue engineering (Colloids and Surfaces B: Biointerfaces79 (2010) 315–325), drug delivery (International Journal of Pharmaceuticals 430 (2012) 335–341), etc. widespread attention in the field.

However, an important problem in the electrospinning of protein polymers is that the prepared silk does not have water resistance, because most proteins are very soluble in water. The solution of the prior art is to subject the protein-containing electrospun yarn to a cross-linking reaction after the electrospun yarn is formed, so as to prevent the protein in the silk from dissolving in water. For example, the steam cross-linking method of glutaraldehyde aqueous solution (ActaBiomaterialia6 (2010) 372-38) reported in the literature, the soaking cross-linking method of glutaraldehyde methanol solution (Journal of Biomedical Material Research A99 (2011) 395) and so on. These crosslinking methods can well inhibit the dissolution of proteins in water during electrospinning, but at the same time, they also lead to the melting of proteins that have formed nanofilament structures during the process of crosslinking, thereby destroying The nanostructure formed by electrospinning increases the diameter of the filament and reduces the specific surface area of the material (Colloids and Surfaces B: Biointerfaces 79 (2010) 315–325; International Journal of Pharmaceuticals 430 (2012) 335–341).

Contents of the invention

In order to solve the above-mentioned technical problems, the present invention provides a method of electrospinning for preparing protein-containing fibers, which is characterized in that: the protein-containing polymer solution is electrospun in an environment containing cross-linking agent and water vapor; The crosslinking agent is a C2-C10 aldehyde compound containing two aldehyde groups.

Electrospinning device is adopted, including electrospinning chamber and spinneret;

The atmosphere of the crosslinking agent and water is obtained by any one of the following two ways:

(1) The protein-containing polymer solution is brought into the electrospinning chamber through the spinneret, and the crosslinking agent aqueous solution is placed in the electrospinning chamber, and the volume concentration of the crosslinking agent is 20% to 80%;

(2) A sleeve-type spinneret containing two coaxial capillaries is used. The crosslinking agent aqueous solution is brought into the electrospinning chamber through the gap between the inner and outer capillaries, and the protein-containing polymer solution is brought into the electrospinning chamber through the inner capillary. ; wherein the volume concentration of the crosslinking agent aqueous solution is 5% to 20%.

When the method of placing the crosslinking agent aqueous solution in the electrospinning chamber is adopted, the vapor pressure of the crosslinking agent in the electrospinning chamber is greater than or equal to 20Pa, and the vapor pressure of water is greater than or equal to 2400Pa at the beginning of the electrospinning process.

When the method of placing the crosslinking agent aqueous solution in the electrospinning chamber is adopted, the vapor pressure of the crosslinking agent in the electrospinning chamber is 20-30000Pa, and the vapor pressure of water is 2400-6000Pa at the beginning of the electrospinning process.

The casing type spinneret is adopted, and the flow ratio of the aqueous solution of the crosslinking agent in the outer layer and the protein-containing polymer solution in the inner layer in the spinneret is between 0.5 and 2.

The polymer component in the protein-containing polymer solution is protein, or a mixture of protein and other non-protein polymers; the mass concentration of protein in the solution is 0.5% to 50%.

The molecular weight of the protein in the protein-containing polymer solution is greater than 10000;

The other non-protein polymers are polymers with a molecular weight greater than 5000 that can be dissolved in the same solvent as proteins;

The solvent of the protein-containing polymer solution is one or more than two organic or inorganic solvents capable of dissolving proteins.

The solvent of the protein-containing polymer solution is one or more of water, acid solution, alkali solution, ethanol, methanol, isopropanol, and hexafluoroisopropanol;

Said other non-protein polymers are polyaniline, polyurethane, poly(ε-caprolactone), chitosan, polyisopropylacrylamide, polyglycolide, polyethylene glycol, polystyrene , polyvinyl chloride, polyacrylonitrile, polylactic acid, polymethyl methacrylate;

The protein is collagen, silk fibroin, serum albumin, zein.

When adopting the method of placing the crosslinking agent aqueous solution in the electrospinning chamber, a sleeve-type spinneret containing two coaxial capillaries can also be used. The protein-containing polymer is divided into two parts and introduced into the electrospinning chamber, and a part of the solution is passed through the electrospinning chamber. The gap between the inner and outer capillaries is brought into the electrospinning chamber, and another part of the solution is brought into the electrospinning chamber through the inner capillary;

Part of the solution may be a protein-free polymer solution that cannot be dissolved in the same solvent as the protein.

The C2-C10 aldehyde compound containing two aldehyde groups is one or both of subaldehyde, pimellaldehyde, adipaldehyde, glutaraldehyde, glyoxal, malondialdehyde or succinaldehyde above.

The present invention has the following advantages:

1. Protein-containing electrospinning with strong water resistance and large specific surface area can be obtained, which avoids the influence of cross-linking reaction on the morphology of electrospinning, and does not cause the fusion between electrospinning fibers after cross-linking to reduce the material The phenomenon of specific surface area.

2. The method is simple and easy to operate, and has a wide range of applications. It can be used for electrospinning of protein solutions, protein-non-protein polymer mixtures, and protein-containing polymer solutions doped with functional materials (such as nanomaterials).

Description of drawings

Figure 1 is a transmission electron micrograph of collagen-polystyrene core-shell structure electrospinning.

In the figure: A-1 and A-2 are the electron micrographs of the core-shell collagen that have not been soaked in water; B-1 and B-2 are the electron micrographs of the core-shell collagen soaked in water for one day; A-2 and B -2 is a partial enlarged view.

Figure 2 is a scanning electron micrograph of pure collagen electrospinning.

detailed description

Implementation mode one:

Place the cross-linking agent aqueous solution in the airtight electrospinning chamber, and after a period of time, the cross-linking agent and water atmosphere will be formed in the electrospinning chamber. When the vapor pressure of the crosslinking agent in the electrospinning chamber is greater than or equal to 20Pa, and the vapor pressure of water is greater than or equal to 2400Pa, electrospinning can be started.

Implementation mode two:

A spinneret containing two coaxial capillaries is used, the capillary in the outer layer and the inner layer pipe is filled with a cross-linking agent aqueous solution, and the inner capillary is filled with a protein-containing polymer solution. During electrospinning, the flow rate of the outer layer and the inner layer solution is adjusted so that the flow rate ratio of the crosslinking agent aqueous solution and the protein-containing polymer solution is between 0.5-2, and stable protein-containing electrospinning can be obtained.

Example

Example 1: Collagen-polystyrene core-shell structure electrospinning by electrospinning by placing glutaraldehyde aqueous solution in the electrospinning chamber

1) Configuration of the polymer solution: the inner layer polymer solution is 5% polystyrene with a final mass concentration dissolved in chloroform-hexafluoroisopropanol mixed solution (v:v=7:3) (A solution), and the outer layer is polymerized The final solution was 5% collagen dissolved in hexafluoroisopropanol (solution B).

2) Construction of the electrospinning device: the electrospinning spinneret is a coaxial structure, the inner and outer pipes are pumped into the solution B, and the inner pipe is pumped into the A solution. Lay an aluminum foil directly under the spinneret as a receiving pole, and add high pressure between the receiving pole and the coaxial spinneret. The spinneret and the receiving electrode are sealed in the electrospinning chamber, which is a plexiglass chamber. The temperature is room temperature (25°C).

3) Electrospinning process: first place an aqueous solution of glutaraldehyde (glutaraldehyde volume concentration 25%) in the electrospinning chamber for more than one hour. Then pump liquid A into the inner layer of the coaxial spinneret at a speed of 0.5mL/min, and at the same time pump liquid B into the inner and outer gaps of the coaxial spinneret at a speed of 0.2mL/min. Add 20000V DC high voltage between the head and the receiving pole. The receiving pole collects the electrospinning.

4) Post-processing of electrospinning: Divide electrospinning into two batches, one batch is directly subjected to transmission electron microscope test (results are shown in Figure 1 (A-1) and (A-2)), the other batch is soaked in water for one day and then The transmission electron microscopy test was carried out (the results are shown in Figure 1 (B-1) and (B-2)).

5) Analysis results: It can be seen from Figure 1 that the prepared electrospun yarn has no fusion phenomenon between silk and silk, and has good water resistance in water at the same time. At the same time, it was observed under the transmission electron microscope that the shell layer of collagen was basically unchanged. It shows that the degree of cross-linking of the outer layer of collagen has a good ability to resist water dissolution.

Example 2: The method of using the coaxial spinneret to pump the cross-linking agent aqueous solution to make collagen electrospinning

(1) Configuration of the polymer solution: the polymer solution of the inner layer is 10% collagen dissolved in hexafluoroisopropanol (solution A). The outer layer crosslinking agent solution is a 10% volume concentration glutaraldehyde-water solution (B solution).

(2) The electrospinning device is as described in Example 1.

(3) Electrospinning process: Pump liquid A into the inner layer of the coaxial spinneret at a speed of 0.5mL/min. When the flow rate is stable, add 10000V DC high voltage between the spinneret and the receiving electrode. After the voltage is stabilized, liquid B is pumped into the inner and outer gaps of the coaxial spinneret at a speed of (0.25-1.0) mL/min, and the electrospinning is collected by the receiving electrode.

(4) Characterization of the electrospun membrane: the membrane was subjected to scanning electron microscopy (results shown in Figure 2).

(5) Analysis results: It can be seen from Figure 2 that the prepared electrospun yarn has no fusion phenomenon between filaments.

Example 3: Making zein electrospinning by placing a succinic dialdehyde aqueous solution in the electrospinning chamber

(1) Configuration of the polymer solution: the polymer solution is 20% zein in mass concentration dissolved in 70% volume concentration of acetic acid aqueous solution (A solution).

(2) Construction of the electrospinning device: as described in Example 1, wherein the electrospinning spinneret was changed to a single capillary (inner diameter: 0.3 mm).

(3) Electrospinning process: place a succinic dialdehyde aqueous solution (50% volume concentration of succinic dialdehyde) in the electrospinning chamber for 30 minutes. Pump liquid A into the capillary at a speed of 1.0mL/min. When the flow rate is stable, a 15000V DC high voltage is applied between the spinneret and the receiving electrode. The receiving pole collects the electrospinning.

Example 4: Collagen electrospinning doped with carbon nanotubes is produced by using a coaxial spinneret to pump an aqueous solution of a crosslinking agent

(1) Configuration of the polymer solution: the polymer solution of the inner layer is collagen with a mass concentration of 20% dissolved in an aqueous solution of acetic acid with a volume concentration of 40% (solution A). After dissolving, add 0.1% mass concentration of sodium dodecylsulfonate and 2% mass concentration of multi-walled carbon nanotubes to solution A, and disperse the multi-walled carbon nanotubes in aqueous acetic acid solution by ultrasonic. The outer layer crosslinking agent solution is 10% glutaraldehyde-water solution (B solution).

(2) The electrospinning device is as described in Example 1.

(3) Electrospinning process: Pump liquid A into the inner layer of the coaxial spinneret at a speed of 0.5mL/min. When the flow rate is stable, add 10000V DC high voltage between the spinneret and the receiving electrode. After the voltage is stabilized, liquid B is pumped into the inner and outer gaps of the coaxial spinneret at a speed of (0.25-1.0) mL/min, and the electrospinning is collected by the receiving electrode.

Claims (10)

1. prepare the electrospinning process containing azelon, it is characterized in that: protein-contg polymer solution is carried out electrospinning in the environment of the steam containing crosslinking agent and water; Described crosslinking agent is the aldehyde compound containing two aldehyde radicals of C2-C10.

2. according to method described in claim 1, it is characterized in that: adopt electrostatic spinning apparatus, comprise electrospinning room, spinning head;

The atmosphere of described crosslinking agent and water is obtained by any one in two kinds of modes below:

(1) protein-contg polymer solution brings electrospinning indoor into through spinning head, the indoor placement cross-linking agent aqueous solution of electrospinning, and crosslinking agent volumetric concentration is 20% ~ 80%;

(2) adopt the bushing type spinning head containing two coaxial capillaries, cross-linking agent aqueous solution brings electrospinning indoor into through the intercapillary space of ectonexine, and protein-contg polymer solution brings electrospinning indoor into through internal layer capillary; Wherein the volumetric concentration of cross-linking agent aqueous solution is 5% ~ 20%.

3. in accordance with the method for claim 2, it is characterized in that: adopt when the mode of the indoor placement cross-linking agent aqueous solution of electrospinning, when electrostatic spinning process starts, the vapour pressure of the indoor crosslinking agent of electrospinning is more than or equal to 20Pa, and the vapour pressure of water is more than or equal to 2400Pa.

4. according to the method described in Claims 2 or 3, it is characterized in that: adopt when the mode of the indoor placement cross-linking agent aqueous solution of electrospinning, the vapour pressure 20-30000Pa of the indoor crosslinking agent of electrospinning, the vapour pressure 2400-6000Pa of water when electrostatic spinning process starts.

5. in accordance with the method for claim 2, it is characterized in that: adopt bushing type spinning head, outer cross-linking agent aqueous solution and internal layer contain the flow-rate ratio of protein polymer solution in spinning head between 0.5 ~ 2.

6. according to the method described in claim 1,2 or 5, it is characterized in that: in described protein-contg polymer solution, component of polymer is the mixture of protein or protein and other non-protein polymer; The mass concentration of Proteins In Aqueous Solutions is 0.5% ~ 50%.

7. in accordance with the method for claim 6, it is characterized in that:

The molecular weight of the protein in described protein-contg polymer solution is greater than 10000;

Other non-protein polymer described is that molecular weight is greater than 5000, can be dissolved in the polymer of same solvent with protein;

The solvent of described protein-contg polymer solution is one or two or more kinds organic or inorganic solvent of energy solubilising protein.

8. in accordance with the method for claim 7, it is characterized in that:

The solvent of protein-contg polymer solution is water, the aqueous solution of pH2-9, ethanol, methyl alcohol, isopropyl alcohol, one or two or more kinds in hexafluoroisopropanol;

Other non-protein polymer described is one or two or more kinds in PLA, polymethacrylates, poly-(6-caprolactone), polyaniline, polyurethane, shitosan, PNIPAM, polyglycolide, polyethylene glycol;

Described protein is collagen, fibroin albumen, seralbumin, one or two or more kinds in zein.

9. in accordance with the method for claim 2, it is characterized in that: adopt when the mode of the indoor placement cross-linking agent aqueous solution of electrospinning, also the bushing type spinning head containing two coaxial capillaries can be adopted, described protein-contg polymer is divided into two part solution and introduces electrospinning indoor, part solution brings electrospinning indoor into through the intercapillary space of ectonexine, and another part solution brings electrospinning indoor into through internal layer capillary;

Wherein a part of solution can be not protein-contg and cannot be dissolved in the polymer solution of same solvent with albumen;

Described not protein-contg and be one or two or more kinds in polystyrene, polyvinyl chloride, polyacrylonitrile with the polymer that albumen cannot be dissolved in same solvent.

10. in accordance with the method for claim 1, it is characterized in that: described C2-C10's is suberic aldehyde containing the aldehyde compound of two aldehyde radicals, heptan dialdehyde, hexandial, glutaraldehyde, glyoxal, one or two or more kinds in malonaldehyde or butanedial.