A kind of preparation method of water-stable γ-polyglutamic acid nanofiber

A technology of polyglutamic acid and nanofibers, applied in fiber treatment, chemical post-treatment of synthetic polymer artificial filaments, single-component polypeptide artificial filaments, etc., can solve the problem of inability to obtain water-stable γ-PGA nanofibers, There are no problems such as γ-PGA nanofiber crosslinking, adverse effects on operators and the environment, and achieve the effects of mild process conditions, good biocompatibility, and easy process operation

Inactive Publication Date: 2011-12-14
DONGHUA UNIV
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Problems solved by technology

Yoshida et al reported the successful preparation of electrospun γ-PGA nanofibers by using TFA as the solvent of the spinning solution, and cross-linked the prepared γ-PGA nanofibers by cystamine dihydrochloride, and obtained the γ-PGA nanofibers under physiological conditions. The γ-PGA-SS nanofiber non-woven fabric that can stably exist under the environment, but the solvent used in this report is pure TFA, which has a strong irritant, and improper operation may have adverse effects on the operator and the environment, and the report does not Control of the size of γ-PGA nanofibers is not achieved by controlling the composition of the spinning solution
In another report, Lee et al. prepared γ-PGA (molecular weight 2000kDa) nanofibers by electrospinning with the aid of polyethylene glycol and Triton X-10, but this report did not pass proper The method cross-links the obtained γ-PGA nanofibers, so water-stable γ-PGA nanofibers cannot be obtained

Method used

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  • A kind of preparation method of water-stable γ-polyglutamic acid nanofiber
  • A kind of preparation method of water-stable γ-polyglutamic acid nanofiber
  • A kind of preparation method of water-stable γ-polyglutamic acid nanofiber

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Embodiment 1

[0035] Take 0.8 grams of γ-PGA powder respectively, dissolve them in 5%, 8% and 10% TFA aqueous solution, stir magnetically for 8 hours to fully dissolve the γ-PGA powder, and prepare three kinds of γ-PGA with a mass percentage concentration of 16%. The solution was stored in a refrigerator at 4°C for later use. When spinning, inject the above three spinning liquids into the syringe pump respectively, the electrospinning process parameters are spinning voltage 32kV, receiving distance 40cm, flow rate 0.3mL / h, humidity not higher than 40%, spinning needle No. 17 Stainless steel needle (1.07mm inner diameter, 1.47mm outer diameter). The aluminum foil collected with the above three different nanofibers was dried in a vacuum desiccator for 24 hours, and then the above aluminum foil was cut to a suitable size and placed in absolute ethanol to peel the fibers from the aluminum foil. SEM observation showed (as shown in FIG. 2 ) that the fibers obtained above had regular morphology a...

Embodiment 2

[0037]Weigh 50 mg of each of the above three different nanofibers and immerse them in 25 mL of anhydrous methanol respectively. Each weighed 750mg of EDC activator, and dissolved it in the above 25mL of anhydrous methanol to activate the carboxyl group on the γ-PGA polymer chain. The activation time was 3 hours, and the activation condition was shaking at room temperature. Then, 872 mg of cystamine dihydrochloride were added to each of them, and they were also reacted in a shaker at room temperature for 72 hours. After the reaction, the fiber was taken out and washed with deionized water for 3 times, and the washing time was 5 minutes each time. After freeze-drying, gamma-PGA nanofibers with good water stability are obtained. In order to evaluate the water solubility of γ-PGA nanofibers cross-linked with cystamine dihydrochloride, a small amount of cross-linked fibers were soaked in PBS buffer for 7 days, and the morphology of the fibers after soaking for 7 days was observed ...

Embodiment 3

[0039] γ-PGA nanofibers with a diameter of 274±51 nm were selected, cut into small pieces of 2 cm×2 cm, spread on a cover glass with a diameter of 1.8 cm and placed in a 24-well cell culture plate. The fibers spread on the cover glass were sterilized by ultraviolet light and soaked in DMEM high-sugar medium for 12h, then supplemented with 400 μL medium per well and inoculated 2×10 4 L929 cells. Different culture time was set for each culture plate (1, 3, 5 and 7 days and 1, 2, 4 and 8 hours, 8 culture plates in total). At the end of each culture period, 40 μL of MTT was added to the culture wells, and the culture was continued for 4 hours, so that the living cells could fully react with MTT to form MTT formazan crystals. Discard the medium in each well, add 400 μL DMSO, completely dissolve the MTT formazan crystals and record the OD value at 570 nm, and analyze the adhesion or proliferation of cells on the surface of γ-PGA nanofibers under different culture times. The effect...

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Abstract

The invention relates to a preparation method of water stability gamma-polyglutamic acid nanometer fibers. The preparation method comprises the steps of: (1) preparing trifluoroacetic acid water solution of which the mass percentage concentration is 5%-10%, dissolving gamma-polyglutamic acid in the trifluoroacetic acid solution and stirring the solution for 6-8 hours to obtain gamma-polyglutamic acid solution; (2) spinning the gamma-polyglutamic acid solution via static spinning to obtain gamma-polyglutamic acid nanometer fibers; and (3) dissolving the gamma-polyglutamic acid nanometer fibersand EDC (ethylene dichloride) in methanol to have a reaction at room temperature for 3-5 hours, adding cystamine dihydrochlorid to have a reaction for 48-72 hours after the reaction is ended, rinsing, freezing and drying the products after the reaction of cystamine dihydrochlorid is ended, thus obtaining gamma-polyglutamic acid nanometer fibers with good water stability. The preparation method ofthe water stability gamma-polyglutamic acid nanometer fibers is simple and convenient in operation and has moderate technique conditions; the raw materials are easy to obtain, and the prepared gamma-polyglutamic acid nanometer fibers have good biocompatibility so the gamma-polyglutamic acid nanometer fibers have latent application merits in the field of the histological engineering bracket.

Description

technical field [0001] The invention belongs to the field of preparation of gamma-polyglutamic acid nanofibers, in particular to a preparation method of water-stable gamma-polyglutamic acid nanofibers. Background technique [0002] Electrospinning (electrostatic spinning) refers to a new spinning method that uses electrostatic field force to make charged polymer solution or melt jet and draw in an electrostatic field to form micro-nano-sized fibers. Electrospinning technology has a series of advantages: firstly, the diameter range of fibers prepared by electrospinning technology is generally between nanometers and microns, which is much smaller than the diameter of fibers prepared by conventional methods, and the obtained fibers have high porosity, It has the advantages of large specific surface area, high fiber fineness and uniformity. These excellent properties that cannot be obtained by traditional spinning methods endow electrospun fibers with a wide range of applicatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): D01F11/08D01F6/68D01D5/00
Inventor 史向阳王世革
Owner DONGHUA UNIV
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