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Method for preparing silk fibroin/hyaluronic acid double-network hydrogel capable of realizing three-dimensional loading of cells

A technology of hyaluronic acid and silk fibroin, which is applied in the fields of medical science and prostheses, can solve the problems that cannot meet the requirements of three-dimensional cartilage construction, difficult three-dimensional loading of cells, toxic cross-linking agents, etc., and achieve adhesion, proliferation and In-situ three-dimensional loading, green and environmentally friendly gelation method, stable physical and chemical properties

Active Publication Date: 2019-05-28
CHONGQING UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the current methods for preparing double network hydrogels use toxic cross-linking agents, or the gelation time is too long, it is difficult to achieve three-dimensional loading of cells in vitro, and cannot meet the requirements of three-dimensional construction of tissues such as cartilage in vitro

Method used

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  • Method for preparing silk fibroin/hyaluronic acid double-network hydrogel capable of realizing three-dimensional loading of cells

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Experimental program
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Effect test

Embodiment 1

[0034] 1. A preparation method of silk fibroin / hyaluronic acid double network hydrogel capable of realizing three-dimensional loading of cells, comprising the following steps:

[0035] 1) Sterilization of materials: using a high temperature and high pressure method, the silk fibroin solution and the modified hyaluronic acid solution are sterilized separately under a high temperature and high pressure environment of 120°C and 0.25 MPa, and the sterilization time is 20 minutes.

[0036] 2) Mix the silk fibroin solution with a mass volume fraction of 8% and the modified hyaluronic acid solution with a mass volume fraction of 2% at a mass ratio of 10:0 to form a prepolymer solution, and add photoinitiator to the prepolymer solution Agent 2-hydroxy-4′-(2-hydroxyethoxy)-2-methylpropiophenone, wherein the mass-volume ratio (g / ml) of the photoinitiator and the prepolymer solution is 0.5%;

[0037] 3) Ultrasound the solution obtained in step 2) for 15s, and the solution is fluid and transpare...

Embodiment 2

[0040] 1. A preparation method of silk fibroin / hyaluronic acid double network hydrogel capable of realizing three-dimensional loading of cells, comprising the following steps:

[0041] 1) Sterilization of materials: The high temperature and high pressure method is used in advance, and the silk fibroin solution and the modified hyaluronic acid solution are sterilized for 20 minutes under a high temperature and high pressure environment of 120°C and 0.25Mpa;

[0042] 2) Mix the silk fibroin solution with a mass volume fraction of 8% and the methacrylic anhydride-modified hyaluronic acid solution with a mass volume fraction of 2% at a mass ratio of 9.5:0.5 to form a prepolymer solution. Add the photoinitiator 2-hydroxy-4'-(2-hydroxyethoxy)-2-methylpropiophenone, wherein the mass volume ratio (g / ml) of the photoinitiator to the prepolymer solution is 0.5% ;

[0043] 3) Ultrasound the solution obtained in step 2) for 13 seconds, and the solution is fluid and transparent after ultrasound;...

Embodiment 3

[0046] 1. A preparation method of silk fibroin / hyaluronic acid double network hydrogel capable of realizing cell encapsulation, including the following steps:

[0047] 1) Sterilization of materials: using a high temperature and high pressure method, the silk fibroin solution and the modified hyaluronic acid solution are sterilized separately under a high temperature and high pressure environment of 120°C and 0.25 MPa, and the sterilization time is 20 minutes.

[0048] 2) Mix the silk fibroin solution with a mass volume fraction of 8% and the modified hyaluronic acid solution with a mass volume fraction of 2% at a mass ratio of 9:1 to form a prepolymer solution, and add photoinitiator to the prepolymer solution Agent 2-hydroxy-4′-(2-hydroxyethoxy)-2-methylpropiophenone, the mass-volume ratio (g / ml) of photoinitiator and prepolymer solution is 0.5%;

[0049] 3) Ultrasound the solution obtained in step 2) for 12s, the solution is fluid and transparent after ultrasound;

[0050] 4) After ...

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Abstract

The invention discloses a method for preparing silk fibroin / hyaluronic acid double-network hydrogel capable of realizing three-dimensional loading of cells. The method comprises the following steps that (1) materials are sterilized; (2) a silk fibroin solution and a modified hyaluronic acid solution are mixed to form a prepolymer solution, and a photoinitiator is added to the prepolymer solution;(3) the prepolymer solution obtained in the step (2) is subjected to ultrasonic treatment, and after the ultrasonic treatment, the solution has fluidity and is translucent; (4) the solution obtained after being treated in the step (3) is mixed with the cells to obtain a cell prepolymerization solution, and the cell prepolymerization solution is further added to a cell culture plate; and (5) the culture plate obtained in the step (4) is placed under ultraviolet light and subjected to photo-crosslinking into gel, and then placed in an incubator at 37 DEG C until the prepolymerization solution becomes milky white, and the silk fibroin / hyaluronic acid double-network hydrogel is obtained. The method can realize the three-dimensional culture of the cells in the high-strength and high-tough double-network hydrogel, and is beneficial to the three-dimensional construction of the load-bearing soft tissue such as cartilage in vitro.

Description

Technical field [0001] The invention belongs to the field of tissue engineering biomaterials, and specifically relates to a preparation method of a silk fibroin / hyaluronic acid double network hydrogel capable of realizing three-dimensional loading of cells. Background technique [0002] The hydrogel material is a polymer with a three-dimensional network system with high water absorption and water retention, and a soft texture, which is similar to the texture of living soft tissue; because its three-dimensional network structure is similar to natural extracellular matrix, it is beneficial to the growth of seed cells. Survival; it can be minimally invasively injected into the body to form a semi-solid colloid in the liquid state to reduce implantation trauma and other unique advantages. It has been widely used in the research of scaffold materials in tissue engineering. However, hydrogel has disadvantages such as poor mechanical properties and excessive degradation speed, which mak...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J3/075C08J3/24C08L89/00C08L5/08A61L27/20A61L27/22A61L27/52
Inventor 肖文谦曲晓航李加乐谭云飞李波
Owner CHONGQING UNIVERSITY OF SCIENCE AND TECHNOLOGY
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