Preparation method of silk fibroin nanosphere

A silk fibroin and nanosphere technology, applied in the field of biomedicine, can solve the problems of difficult bioactive molecule loading, inactivation of bioactive molecules, loss of biomolecular activity, etc., to maintain biocompatibility, high yield, and size. control effect

Active Publication Date: 2012-04-18
广东圣蜜生物科技有限公司
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

In the technical solutions disclosed in the above-mentioned documents, it is necessary to use denaturants to make silk fibroin insoluble in water. Due to the use of denaturants, it may cause the reduction of biocompatibility and the inactivation of other bioactive molecules, so it is difficult to use Loading of bioactive molecules
[0005] The document "Simple preparation and characteristics of silk fibroin microsphere" ([J]European Polymer Journal 39 (2003) 1195–1199) reported the preparation of silk fibroin microspheres with an average diameter of 2 microns by spray drying, and the microspheres were irregular in shape , have experienced a higher temperature treatment of 85 ° C, which may also caus...

Method used

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  • Preparation method of silk fibroin nanosphere
  • Preparation method of silk fibroin nanosphere

Examples

Experimental program
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Embodiment 1

[0033] Put 100 g of silkworm raw silk into 5 L of 0.05% Na 2 CO 3 In aqueous solution, at 98-100 o C treatment for 30 minutes, repeated 3 times to degumming silk, fully washed and dried to obtain pure silk fiber. Add pure silk fiber to the ternary solution (by CaCl 2 / water / ethanol molar ratio is 1:8:2) solution, at 70 o C stirring and dissolving into a mixed solution of silk fibroin. Put the obtained silk fibroin mixed solution into a dialysis bag and dialyze with deionized water for 4 days to remove CaCl 2、 Ethanol and other impurities are removed to obtain a pure silk fibroin solution.

[0034] Using cyclohexane as a solvent, add emulsifier Span, the amount of Span added is 1%wt of cyclohexane, stir and dissolve to obtain a cyclohexane / Span mixed solution.

[0035] Adjust the mass concentration of the silk fibroin solution to 3%, and evenly add polyol ethylene glycol with 10% silk fibroin mass into the silk fibroin solution.

[0036] Add the silk fibroin / polyol mixed...

Embodiment 2

[0044] Put 0.1 kg of raw silk into 3 liters of sodium carbonate aqueous solution with a mass concentration of 0.15%, treat it at 98-100° C. for 2 hours, degumming the raw silk, and obtain pure silk fiber after fully washing. The dried pure silk fiber is heated and dissolved at 60±5° C. with 1 liter of lithium bromide solution with a concentration of 9.0 mol / liter to obtain a silk fibroin mixed solution. The cellulose membrane is used as a dialysis material, and the obtained silk fibroin mixed solution is dialyzed with deionized water for 3 days to remove impurities such as lithium bromide to obtain a pure silk fibroin solution.

[0045] Cyclohexane is used as a solvent, and the emulsifier Span is added in an amount of 0.75%wt of cyclohexane, and stirred and dissolved to obtain a cyclohexane / Span mixed solution.

[0046] Adjust the concentration of silk fibroin solution to 3%, and add glycerol with 30% silk fibroin mass evenly into the silk fibroin solution.

[0047] Add the s...

Embodiment 3

[0055] Put 200 grams of waste 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 silk fibroin after fully washing. The dried pure silk fibroin is heated and dissolved in 1 liter of calcium chloride, water and ethanol solution with a molar ratio of 1:8:2 at 75±5°C to obtain a silk fibroin mixed solution. The cellulose membrane is used as a dialysis material, and the obtained silk fibroin mixed solution is dialyzed with deionized water for 3 days to obtain a pure silk fibroin solution.

[0056] Using 80 ml of cyclohexane as a solvent, add 0.4 g of emulsifier Span, stir and dissolve to obtain a cyclohexane / Span mixed solution.

[0057] Adjust the concentration of silk fibroin solution to 3.2%, and add glycerol with 30% silk fibroin mass evenly into the silk fibroin solution.

[0058] Add the silk fibroin / polyol mixed solution drop by drop into the cyclohex...

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Abstract

The invention discloses a preparation method of silk fibroin nanospheres, which comprises the following steps: dispersing a silk fibroin solution into nanometer liquid drops by an inverse emulsion method, adding polyols under ultrasonic oscillation to promote silk fibroin in the nanometer liquid drops to form a structure insoluble in water, adding high water-absorbing resins to dry and cure the silk fibroin in a microemulsion state so as to obtain silk fibroin nanospheres which are insoluble in water. The silk fibroin nanospheres have good biocompatibility, are nontoxic and harmless for human body, and have no immunogenicity; the preparation process adopts mild technology with a normal temperature, a normal pressure, and no chemical cross-linking agents; therefore, the product can be used as a carrier with biologically-active substances to load enzymes, nucleic acids, polypeptides, protein drugs, and the like; the obtained silk fibroin spheres have small particle sizes, good dispersibility, do not adhere, and can be prepared into various cosmetics, skin care products, sunscreen creams and the like; the silk fibroin spheres have small particles which can run freely in blood, thus can be used as injections and have wide application in fields of disease diagnosis and treatment.

Description

technical field [0001] The invention relates to a method for preparing organic polymer nanospheres, in particular to a method for preparing nanoparticles by using silk fibroin as a raw material, and belongs to the technical field of biomedicine. 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 like. Microspheres made of biod...

Claims

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Application Information

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IPC IPC(8): C08L89/00C08J3/05A61K8/64A61K47/42A61K9/14A61Q17/04A61Q1/00A61Q19/00
Inventor 邢铁玲卢神州李明忠张岑岑吕强
Owner 广东圣蜜生物科技有限公司
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