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Wild antheraea pernyi silk fibroin microsphere and preparation method thereof

A technology of tussah silk fibroin and tussah silk, applied in the field of biomedical materials, can solve the problems of insufficient cell recognition and targeting, and difficulty in loading with bioactive molecules, and achieve good biocompatibility and maintain biocompatibility , size controllable effect

Inactive Publication Date: 2013-03-20
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the silk fibroin amino acid sequence does not contain the RGD tripeptide sequence, it is insufficient for cell recognition and targeting
And the tussah silk powder obtained by salting out with saturated ammonium sulfate is also difficult to be used for the loading of bioactive molecules.

Method used

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  • Wild antheraea pernyi silk fibroin microsphere and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Put 100 g tussah raw silk into 5 L Na with a mass concentration of 0.05% 2 CO 3 In the aqueous solution, the treatment is carried out at a temperature of 98-100° C. for 30 minutes, and repeated 4 times to degumming the silk. After washing and drying thoroughly, pure tussah silk fibers are obtained. Add pure tussah silk fibroin fiber into saturated lithium thiocyanate solution, stir and dissolve at 50°C to form tussah silk fibroin protein mixed solution. Put the obtained tussah silk fibroin mixed solution into a dialysis bag and dialyze with deionized water for 4 days to remove impurities such as lithium thiocyanate to obtain a pure tussah silk fibroin protein solution.

[0017] Adjust the concentration of tussah silk fibroin protein to 20 mg / mL, add acetic acid and sodium acetate buffer solution with pH=4.3 at a ratio of 1:1 by volume, place the mixture in a water bath environment at 35 °C, and turn on ultrasonic vibration and stirring at the same time, After 60 minut...

Embodiment 2

[0020] Put 0.1 kg of tussah silk into 3 liters of 0.15% sodium carbonate aqueous solution, treat it at 98-100° C. for 2 hours, degumming the tussah silk, and obtain pure tussah silk fiber after fully washing. The dried pure tussah fibroin fiber is heated and dissolved at 50±5°C with 1 liter of lithium thiocyanate solution with a concentration of 9.3 mol / liter to obtain a tussah silk fibroin protein mixed solution. Using the cellulose membrane as the dialysis material, the obtained tussah silk fibroin protein mixed solution was dialyzed with deionized water for 3 days to remove impurities such as lithium thiocyanate to obtain a pure tussah silk fibroin protein solution.

[0021] Adjust the tussah silk fibroin protein concentration to 40 mg / mL, add pH=4 acetic acid and sodium acetate buffer at a volume ratio of 1:1, place the mixture in a water bath at 30 °C, and turn on ultrasonic vibration and stirring at the same time. After 50 minutes, the tussah silk fibroin microsphere sus...

Embodiment 3

[0024] Put 200 grams of tussah silk into 8 liters of sodium carbonate aqueous solution with a mass concentration of 0.05%, degumming at 98-100° C. for 0.5 hour, repeat the treatment 3 times, and obtain pure tussah silk fiber after fully washing. The tussah silk fibroin protein mixed solution is obtained by heating and dissolving the dried tussah silk fibers with 1 liter of lithium thiocyanate solution with a concentration of 9.3 mol / liter at 55±5° C. The cellulose membrane is used as a dialysis material, and the obtained tussah silk fibroin protein mixed solution is dialyzed with deionized water for 3 days to obtain a pure tussah silk fibroin protein solution.

[0025] Adjust the tussah silk fibroin protein concentration to 10 mg / mL, add pH=3 citric acid and sodium citrate buffer solution at a volume ratio of 1:1, place the mixture in a water bath at 35 °C, and turn on the ultrasonic wave at the same time Shaking and stirring, tussah silk fibroin microsphere suspension was obt...

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Abstract

The invention discloses a wild antheraea pernyi silk fibroin microsphere and a preparation method of the microsphere. The preparation method comprises the steps of degumming and dissolving tussah silk; subjecting the above processed tussah silk to dialysis treatment to obtain a solution of the wild antheraea pernyi silk fibroin; adjusting the concentration of the solution to 10-100mg / ml; adding in a citric acid solution or an acetic acid buffer solution at the temperature of 10-60 DEG C to adjust the pH value to 3-6; subjecting the resulting solution to ultrasonic oscillation and stirring treatment to obtain a suspension of the wild antheraea pernyi silk fibroin microsphere; centrifugalizing, freezing and drying the suspension to obtain the biodegradable wild antheraea pernyi silk fibroin microsphere, the diameter of which is 0.1-10mu m. The preparation method of the invention prepares the wild antheraea pernyi silk fibroin microsphere by means of in-situ self-assembly generation and is simple in process procedures; the prepared microsphere is uniform in size distribution, keeps the RGD sequence of the wild antheraea pernyi silk fibroin, promotes the cell recognition of the microsphere, improves the targeting property and bioavailability of drugs, can function as a carrier with bioactive substance to carry an enzyme drug, a nucleic acid drug, a polypeptide drug, a protein drug and the like, and can be applied to diagnosing and treating diseases and the like.

Description

technical field [0001] The invention relates to a method for preparing organic polymer microspheres, in particular to a microsphere prepared from tussah silk fibroin as a raw material and a preparation method thereof, belonging to the technical field of biomedical materials. Background technique [0002] In recent years, with the development of biotechnology and genetic engineering, more and more biologically active substances such as proteins and enzymes are expected to be used in the treatment of diseases. However, since protein and polypeptide drugs are easily degraded in the intestinal tract after oral administration, the circulation time in the body is short and the biological half-life is short after intravenous injection, which greatly restricts their application. To solve these problems, various protein drug dosage forms have emerged, including hydrogels, nanospheres, microspheres, liposomes of lipoplexes, solid lipid nanoparticles, water-in-oil emulsions, and the li...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J3/12C08L89/00A61K47/42A61K9/16A61K8/64A61Q19/00A61Q17/04
Inventor 卢神州毛丽李贵军邢铁玲
Owner SUZHOU UNIV
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