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Preparation method of composite gel three-dimensional tumor model scaffold

A composite gel, three-dimensional technology, applied in the field of cell biology, can solve the problems of poor pore size uniformity of scaffold materials, unfavorable tumor cell adhesion, complicated preparation process, etc., and achieves shortened preparation time, suitable pore size and uniformity, The effect of simplifying the synthesis step

Inactive Publication Date: 2016-09-28
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, chitosan itself has certain defects: due to the action of lysozyme, chitosan degrades quickly, and it is easy to lose its mechanical properties and cause structural failure.
[0008] The silk fibroin-chitosan composite porous scaffold published in the prior art has relatively low stability and is difficult to meet the needs of practical applications, and the preparation process is cumbersome and the cycle is long. It is necessary to first prepare the three-dimensional scaffold material, and then add the pore agent, the final porous three-dimensional scaffold material is obtained after treatment. The prepared scaffold material has poor pore size uniformity and poor morphology, which is not conducive to the adhesion of tumor cells.

Method used

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  • Preparation method of composite gel three-dimensional tumor model scaffold
  • Preparation method of composite gel three-dimensional tumor model scaffold
  • Preparation method of composite gel three-dimensional tumor model scaffold

Examples

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Embodiment 1

[0052] 1. Preparation of silk fibroin-chitosan composite gel three-dimensional tumor model scaffold

[0053] Take an appropriate amount of chitosan (molecular weight is 8500Da) and put it into a mortar and grind it finely, and dry it in an oven at 50°C, then weigh 1g of the powder, add it to 100mL of 1% glacial acetic acid aqueous solution, stir to dissolve it, and obtain Chitosan solution with a concentration of 0.01 g / mL. Put the chitosan solution in a refrigerator at 4°C overnight to let the air bubbles out of the solution. Put 10mL of silk fibroin solution with a concentration of 150g / L into a plastic container, slowly add 7mL of chitosan solution with a concentration of 0.01g / mL while stirring, then add 96mg of EDC and 57.5mg of NHS, stir well , 5 mL of n-butanol was added dropwise, and after 5 minutes, the solution was gel-like. Stop stirring, put the gel in a refrigerator at 4°C for 3 minutes to remove air bubbles in the gel, then put it in a refrigerator at -20°C for...

Embodiment 2

[0060] Take an appropriate amount of chitosan and grind it into a mortar, dry it in an oven at 50°C, then weigh 1g of the powder, add it to 100mL of 1% glacial acetic acid aqueous solution, stir to dissolve it, and obtain a concentration of 0.01 g / mL chitosan solution. Put the chitosan solution in a refrigerator at 4°C overnight to let the air bubbles out of the solution. Put 10mL of silk fibroin solution with a concentration of 75g / L into a plastic container, slowly add 7mL of chitosan solution with a concentration of 0.01g / mL while stirring, then add 134.4mg of EDC and 80.5mg of NHS, and stir evenly Finally, 4.25 mL of n-butanol was added dropwise for denaturation. After 5 minutes, the solution was gel-like. Stop stirring, put the gel in the refrigerator at 8°C for 3 minutes to remove the air bubbles in the gel, then put it in the refrigerator at -20°C for 20 minutes, and put it in the refrigerator at -80°C for molding , taken out after 24 hours, and freeze-dried to obtain...

Embodiment 3

[0062] Take an appropriate amount of chitosan and grind it into a mortar, dry it in an oven at 50°C, then weigh 1g of the powder, add it to 100mL of 1% glacial acetic acid aqueous solution, stir to dissolve it, and obtain a concentration of 0.01 g / mL chitosan solution. Put the chitosan solution in a refrigerator at 6°C overnight to let the air bubbles out of the solution. Put 10mL of silk fibroin solution with a concentration of 75g / L into a plastic container, slowly add 3mL of chitosan solution with a concentration of 0.01g / mL while stirring, then add 57.6mg of EDC and 34.5mg of NHS, and stir evenly After that, 3.25 mL of n-butanol was added dropwise, and after 5 minutes, the solution was gel-like. Stop stirring, put the gel in a refrigerator at 4°C for 3 minutes to remove air bubbles in the gel, then put it in a refrigerator at -20°C for 20 minutes, and put it in a refrigerator at -80°C for molding , taken out after 24 hours, and freeze-dried to obtain a silk fibroin-chito...

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Abstract

The invention relates to a preparation method of a composite gel three-dimensional tumor model bracket, comprising the following steps: grinding and drying chitosan, and then dissolving it in an aqueous solution of glacial acetic acid to obtain a chitosan solution; Bubbles; under stirring conditions, add the treated chitosan solution to the silk fibroin aqueous solution, and then add a crosslinking agent to obtain a mixed solution; under stirring conditions, add n-butanol to the mixed solution to obtain a gel; Exclude the air bubbles in the gel, shape and freeze-dry to obtain a composite gel three-dimensional tumor model scaffold. In the present invention, silk fibroin is used as the main material, a small amount of chitosan is used as the auxiliary material, and n-butanol is used as the denaturant to construct tumor tissue engineering scaffold materials in an aqueous solution system, and the prepared composite gel three-dimensional tumor model scaffold is both It can retain the material structure basis of the cell microenvironment in vivo, and can simulate more realistic physiological conditions for tumor cell growth.

Description

technical field [0001] The invention relates to the field of cell biology, in particular to a method for preparing a composite gel three-dimensional tumor model scaffold. Background technique [0002] Cancer is a complex disease that depends on the activation of proto-oncogenes, loss-of-function mutations of tumor suppressor genes, and local and systemic factors under environmental influence. The tumor tissue environment is very complex. In addition to tumor cells, there are many other types of cells, such as epithelial cells, fibroblasts, and immune cells. Variations in cancer genes cause tumors to have different manifestations and characteristics in this complex environment. There are complex signal transduction pathways in different types of cells and between cells in tumor cells. In addition, the complex network structure formed by different cell populations will be further affected by the different physical and chemical environments in which tumor cells live. [0003] ...

Claims

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Application Information

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IPC IPC(8): A61L27/48A61L27/52A61L27/54A61L27/50
CPCA61L27/48A61L27/50A61L27/52A61L27/54A61L2300/232A61L2300/252A61L2300/416C08L5/08
Inventor 张学农李继昭周烨娟
Owner SUZHOU UNIV
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