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Three-dimensional silk fibroin scaffold insoluble in water, and preparation and application of three-dimensional silk fibroin scaffold

A protein scaffold and silk fibroin technology, applied in the field of water-insoluble three-dimensional silk fibroin scaffold and its preparation, can solve the problems of uneven pore wall thickness of porous scaffold, high mechanics of silk fibroin scaffold, slow degradation rate, etc. , to achieve the effect of being conducive to mass production, huge application prospects, and good biocompatibility

Inactive Publication Date: 2013-01-02
ZHEJIANG XINGYUE BIOTECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Taking sodium chloride particles as the way of denaturation and pore formation as an example, the formed silk fibroin gel is deposited in the gaps of sodium chloride particles. Due to the inhomogeneity of the gaps, the thickness of the pore wall of the porous scaffold is inhomogeneous, from several Micron to hundreds of microns, the formed silk fibroin scaffold has high mechanical properties and a slow degradation rate, but it is not easy to completely wash away a small amount of sodium chloride particles completely wrapped by silk fibroin in the scaffold

Method used

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  • Three-dimensional silk fibroin scaffold insoluble in water, and preparation and application of three-dimensional silk fibroin scaffold

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 (30% silk fibroin concentration)

[0023](1) Preparation of silk fibroin solution: a) Degumming: Put 100g of silkworm silk (Zhejiang Huazhi Silk Co., Ltd., 5A grade) into 5L of 2M sodium carbonate aqueous solution, bathe in 98°C for 30min, wash with pure water, Repeat this process 3 times to remove the sericin and leave the silk fibroin. Dry the silk fibroin at 50°C to obtain 70g of dried silk fibroin for later use; b) dissolving: the above-mentioned dried silk fibroin The protein was dissolved in 9.3M lithium bromide (LiBr) aqueous solution at a mass volume ratio of 0.2:1, and the silk fibroin was fully dissolved in a water bath at 60°C for 90 minutes to obtain a mixed solution containing silk fibroin and a small amount of insoluble particles; c) Dialysis: the The mixed solution was dialyzed with a regenerated cellulose dialysis bag (molecular weight cut-off 4000 Daltons), dialyzed 12 times in 3 days with sterile pure water 10 times the volume of the mixed so...

Embodiment 2

[0026] Embodiment 2 (silk fibroin solution concentration 5%)

[0027] (1) Preparation of silk fibroin solution: a) Degumming: Put 100g of silkworm silk (Zhejiang Huazhi Silk Co., Ltd., 5A grade) into 5L of 2M sodium carbonate aqueous solution, bathe in 98°C for 30min, wash with pure water, Repeat this process 3 times to remove the sericin and leave the silk fibroin. Dry the silk fibroin at 50°C to obtain 70g of dried silk fibroin for later use; b) dissolving: the above-mentioned dried silk fibroin The protein was dissolved in 9.3M lithium bromide (LiBr) aqueous solution at a mass volume ratio of 0.2:1, and the silk fibroin was fully dissolved in a water bath at 60°C for 90 minutes to obtain a mixed solution containing silk fibroin and a small amount of insoluble particles; c) Dialysis: the The mixed solution was dialyzed with a regenerated cellulose dialysis bag (molecular weight cut-off 4000 Daltons), dialyzed 12 times with sterile pure water of 10 times the volume of the mix...

Embodiment 3

[0029] Embodiment 3 (silk fibroin solution concentration 1%)

[0030] (1) Preparation of silk fibroin solution: a) Degumming: Put 100g of silkworm silk (Zhejiang Huazhi Silk Co., Ltd., 5A grade) into 5L of 2M sodium carbonate aqueous solution, bathe in 98°C for 30min, and wash with deionized water , this process was repeated 3 times to remove the sericin and leave the silk fibroin, and dry the silk fibroin at 50°C to obtain 70g of dried silk fibroin for later use; b) dissolving: dissolve the silk fibroin by mass Dissolve in 9.3M lithium bromide (LiBr) aqueous solution at a volume ratio of 0.20:1, bathe in water at 60°C for 90 minutes until the silk fibroin is fully dissolved, and obtain a mixed solution containing silk fibroin and a small amount of insoluble particles; c) Dialysis: Dilute the mixed solution with The regenerated cellulose dialysis bag (molecular weight cut-off 2000 Dalton) was dialyzed, and the sterile deionized water of 10 times the volume of the mixed solutio...

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Abstract

The invention discloses a three-dimensional silk fibroin scaffold insoluble in water, and preparation and application of the three-dimensional silk fibroin scaffold. A preparation method includes the steps of injecting silk fibroin solution with the mass concentration of 0.3-30% into a mold, and obtaining a three-dimensional silk fibroin scaffold after freeze-drying; performing humid and heat crosslinking for the mold under the condition that the temperature ranges from 50 DEG C to 100 DEG C and the relative humidity ranges from 70% to 100%; and drying, removing the mold and obtaining the three-dimensional silk fibroin scaffold insoluble in water. Silk fibroin is used as a raw material, and biocompatibility is good; a preparation process is non-toxic and environment-friendly, and shape, aperture and porosity parameters of the scaffold are easy to control; the pore wall of the scaffold is nano-scale, the aperture of the scaffold is micron-scale, an extracellular matrix micro-environment is simulated, and the three-dimensional silk fibroin scaffold is used as a three-dimensional cell culture vector or a three-dimensional plasmid transfection vector when used externally; and the three-dimensional silk fibroin scaffold is used in the aspects of cartilage scaffolds, fat scaffolds, bone scaffolds, muscle scaffolds and the like when used in the internal tissue engineering field.

Description

(1) Technical field [0001] The invention relates to a three-dimensional silk fibroin scaffold and a preparation method thereof, in particular to a water-insoluble three-dimensional silk fibroin scaffold, a preparation method and application thereof. (2) Background technology [0002] Silk is one of the strongest natural fibers, consisting of 70-80% silk fibroin and 20-30% sericin. The amino acid sequence of silk fibroin is mainly composed of repeating units of "glycine-alanine-glycine-alanine-glycine-serine" (GAGAGS), in which glycine, alanine and serine account for more than 85% of the total amino acid content . Most of the repeated sequences are located in the crystallization region of silk fibroin, and self-assemble into an anti-parallel β-sheet structure. These structures endow silk fibroin with the properties of good mechanics and slow degradation rate. [0003] Sericin can cause inflammation in the body and needs to be degummed before use. Silk fibroin is composed ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/22A61L27/50C12N5/00C12N15/63
Inventor 陈隆坤其他发明人请求不公开姓名
Owner ZHEJIANG XINGYUE BIOTECH
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