Preparation method of sulfated silk fibroin material

A technology of sulfonated silk fibroin and silk fibroin, which is applied in the field of preparation of sulfonated silk fibroin materials, can solve the problems of low biological activity, low biological activity, small immunogenicity, etc., and achieve high reaction efficiency and good resistance Effect of coagulation and improvement of adhesion

Inactive Publication Date: 2012-12-19
JIANGSU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Natural polymer materials are very similar to biological macromolecules, can be degraded by enzymes in vivo, have better histocompatibility than artificial synthetic materials, and have less immunogenicity, but they also have shortcomings, such as low biological activity
In recent years, silk fibroin has been widely used in tissue engineering of cornea, blood vessel, artificial skin, ligament and bone, slow and controlled drug release, enzyme immobilization and other fields, but its biological activity is low

Method used

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  • Preparation method of sulfated silk fibroin material
  • Preparation method of sulfated silk fibroin material
  • Preparation method of sulfated silk fibroin material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Mix 2mL of 0.4M aqueous solution of p-aminobenzenesulfonic acid with 1mL of 1.6M aqueous solution of p-toluenesulfonic acid, the molar ratio of p-aminobenzenesulfonic acid to p-toluenesulfonic acid is 1:2, and keep the temperature at 5°C.

[0019] Add 1mL of 0.8M sodium nitrite aqueous solution in ice bath to the mixed liquid in step 1, mix well, and maintain the ice bath for 15 min to obtain the azo salt reaction liquid.

[0020] Dissolve 0.8g silk fibroin in citrate buffer solution (0.05M pH 8) to obtain 8mL silk fibroin solution with a concentration of 10% (w / v, g / mL), add 16mL azo salt reaction solution (2mL / 100mg silk fibroin), maintain the pH at 8 with citric acid buffer, react in an ice bath for 20 minutes, use a dialysis bag with a molecular weight cut-off of 3500 for 2-3 days, and freeze-dry to obtain sulfonated silk fibroin.

Embodiment 2

[0022] Mix 10mL 0.08M p-toluenesulfonic acid aqueous solution with 4mL 1.6M p-toluenesulfonic acid aqueous solution, the molar ratio of p-aminobenzenesulfonic acid to p-toluenesulfonic acid is 1:8, and keep the temperature at 0°C.

[0023] Add 1.6mL of 1M sodium nitrite aqueous solution in ice bath to the mixed liquid in step 1, mix well, and maintain the ice bath for 30min to obtain the azo salt reaction liquid.

[0024] Dissolve 0.8g of silk fibroin in citrate buffer solution (0.05M pH 8) to obtain 4mL of silk fibroin solution with a concentration of 20% (w / v, g / mL), add 0.08mL of azo salt to react solution (0.01mL / 100mg silk fibroin), maintain the pH at 10 with citric acid buffer, react in an ice bath for 15 minutes, use a dialysis bag with a molecular weight cut-off of 3500 for 2-3 days, and freeze-dry to obtain sulfonated silk fibroin.

Embodiment 3

[0026] Mix 100mL of 1mM p-toluenesulfonic acid aqueous solution with 40mL of 0.01M p-toluenesulfonic acid aqueous solution and cool in an ice bath.

[0027] Add 0.0017mL of 6M sodium nitrite aqueous solution in ice bath to the mixture in step 1, mix well, and maintain the ice bath for 20min to obtain the azo salt reaction liquid.

[0028] Dissolve 0.1g silk fibroin in citrate buffer solution (0.05M pH 8) to obtain 10mL silk fibroin solution with a concentration of 1% (w / v, g / mL), add 1mL azo salt reaction solution (1 mL / 100 mg silk fibroin), maintain the pH at 9 with citric acid buffer, react in an ice bath for 10 min, dialyze for 2-3 days with a dialysis bag with a molecular weight cut-off of 3500, and freeze-dry to obtain sulfonated silk fibroin.

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Abstract

The invention provides a preparation method of a sulfated silk fibroin material. The method comprises the steps of: a, preparing an azo salt stock solution; b, dropping ice-bath cooled sodium nitrite aqueous solution to a mixed solution in the step a, and carrying out ice-bath uniformly to obtain the azo salt stock solution; and c, dissolving silk fibroin in a citrate buffer solution to obtain a silk fibroin solution, adding the azo salt stock solution, maintaining the pH of the citrate buffer solution within 7.5-10, carrying out ice-bath reaction, and dialyzing, freezing and drying to obtain the sulfated silk fibroin. The product prepared by the invention is not only a material with good anticoagulant performance, but also has less immunogenicity, so that the material is a bioactive material which can improve conglutination, diffusion and proliferation capacity of the silk fibroin, cells and growth factors. Types of biomaterials are enriched, and grounds for tissue engineering application such as vessels, ligaments and bones are provided.

Description

technical field [0001] The invention belongs to the field of biological materials, and in particular relates to a preparation method of a sulfonated silk fibroin material which can be used for ligament, blood vessel and bone tissue engineering materials. Background technique [0002] Biomaterials are a new type of high-tech materials used in the treatment, diagnosis, repair or replacement of human tissues and organs or to enhance their functions. They involve the health of hundreds of millions of people and are necessary to ensure human health. Their successful application has saved the lives of critically ill patients. Life, but also improve the quality of life and health of patients. Natural polymer materials are very similar to biological macromolecules, can be degraded by enzymes in vivo, have better histocompatibility than artificial synthetic materials, and have less immunogenicity, but they also have shortcomings, such as low biological activity. In order to maintain...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08H1/00A61L27/22A61L33/12
Inventor 颜辉李绍群唐玉斌江明珠贾俊强吴琼英
Owner JIANGSU UNIV OF SCI & TECH
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