A preparation method and application of a silk fibroin frozen sponge capable of slow-release exosomes

A silk fibroin and exosome technology, which is applied in the field of preparation of silk fibroin frozen sponge, can solve the problems of too fast release of exosomes, disappearance of biological regulation functions, and inability to realize late regulation and other problems.

Active Publication Date: 2020-12-11
PEKING UNIV THIRD HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the face of large-scale tissue repair and tissue repair with insufficient blood supply, it usually takes several months, which leads to the release of exosomes too fast and cannot be adjusted later;
[0007] 3. After the release is completed, the stent still exists, resulting in the disappearance of its biological regulation function, and can only provide mechanical support
The applicant found in the research that the silk fibroin frozen sponge technology disclosed in the previous patent is not suitable for the sustained release of exosomes, and further development and exploration of the sustained release system is needed

Method used

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  • A preparation method and application of a silk fibroin frozen sponge capable of slow-release exosomes
  • A preparation method and application of a silk fibroin frozen sponge capable of slow-release exosomes
  • A preparation method and application of a silk fibroin frozen sponge capable of slow-release exosomes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] 1) Silk fibroin extraction

[0058] Cut 10 g of silkworm cocoons into small pieces with scissors, put them into 0.02 mol / L anhydrous sodium carbonate solution (2 L) after boiling, stir and dissolve for 30 min while maintaining the boiling state, and take out the white cotton-like solid obtained, Wash once and dry at room temperature. After drying fully, weigh and weigh the corresponding lithium bromide with the mass ratio of silk fibroin: lithium bromide = 1:4, and prepare a 9.3 mol / L lithium bromide aqueous solution (calculate the liquid volume first, then weigh about 60% of the water , ice bath, at the same time slowly add lithium bromide to the stirring water, make up the volume to the calculated value after completely dissolving), and pour the solution into the beaker containing silk fibroin, so that the lithium bromide solution completely covers the protein, then at 60 ℃ Stir for 4 hours, then dialyze the obtained yellow-brown viscous solution, change the water ev...

Embodiment 2

[0066] 1) Silk fibroin extraction

[0067] Cut 15 g of silkworm cocoons into small pieces with scissors, put them into 0.02 mol / L anhydrous sodium carbonate solution (2 L) after boiling, stir and dissolve for 60 min while keeping the boiling state, take out the obtained white cotton-like solid, and Wash once and dry at room temperature. After drying fully, weigh and weigh the corresponding lithium bromide with the mass ratio of silk fibroin: lithium bromide = 1:4, and prepare a 9.3 mol / L lithium bromide aqueous solution (calculate the liquid volume first, then weigh about 60% of the water , ice bath, at the same time slowly add lithium bromide to the stirring water, make up the volume to the calculated value after completely dissolving), and pour the solution into the beaker containing silk fibroin, so that the lithium bromide solution completely covers the protein, then at 60 ℃ Stir for 4 hours, then dialyze the obtained yellow-brown viscous solution, change the water every ...

Embodiment 3

[0074] Example 3 The ability of silk fibroin to freeze into a sponge is affected by ionic strength and protein concentration

[0075] In order to analyze the effect between the preparation ability of the frozen sponge and the ionic strength and protein concentration, silk fibroin solutions with different ionic strength and protein concentration were prepared, and the glass transition temperatures of these batches of protein were compared using differential thermal experiments . It has been found through experiments that the greater the ionic strength, the lower the glass transition temperature of the silk fibroin solution and the poorer the ability to form a sponge. In contrast, the higher the protein concentration, the less the effect of ionic strength (attached figure 1 ).

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Abstract

The invention provides a preparation method and application of a silk fibroin frozen sponge capable of slowly releasing exosome. Compared with traditional methods, according to an embodiment of the invention, long-term storage of the silk fibroin frozen sponge in a frozen environment is realized; on the premise of ensuring that a scaffold structure is not damaged, and a release rate is reduced, scaffold degradation and exosome release are synchronously conducted; and the exosome release rate can be regulated and controlled only by regulating and controlling the degradation rate of the silk fibroin frozen sponge, so the silk fibroin frozen sponge has wide market prospects.

Description

technical field [0001] The invention relates to the field of biological scaffolds, in particular to a preparation method and application of a silk fibroin frozen sponge of exosomes. Background technique [0002] Exosomes are lipid bilayer vesicles that are released from endosomes to the outside of cells through various transport mechanisms. There have been a lot of studies exploring the structure and function of exosomes. The diameter of exosomes is between 50-100 nm, and the shape is mostly spherical. After inward budding through the endosomal membrane to form multivesicular bodies, the multivesicular bodies fuse with the plasma membrane, thereby releasing exosomes into the extracellular environment. Exosomes are important nanovesicles that regulate intercellular communication. The molecular components they carry include lipids (prostaglandins, phosphokinase C / D, etc.), proteins (transmembrane proteins, lipid-anchored membrane proteins, external peripheral membrane Protei...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08J9/28C08J3/24A61L27/22A61L27/38A61L27/54A61L27/56A61L27/58C07K14/435C12N5/0735C12N5/077C12N5/0775C08L89/00
CPCA61L27/227A61L27/3804A61L27/3834A61L27/3895A61L27/54A61L27/56A61L27/58A61L2300/30A61L2300/412A61L2300/604C07K14/43586C08J3/24C08J9/28C08J2201/0482C08J2389/00C12N5/0606C12N5/0657C12N5/0665C12N2509/00C12N2509/10C08L89/00
Inventor 敖英芳孙牧旸余慧镭胡晓青
Owner PEKING UNIV THIRD HOSPITAL
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