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Injected chemical crosslinking protein/polypeptide hydrogel and preparation method thereof

A chemical cross-linking and protein technology, applied in the field of chemical cross-linking protein/polypeptide hydrogel and its preparation, can solve the problems of short gel time, mechanical strength, limitation of photo-cross-linking hydrogel, cytotoxicity, etc. The effect of stable 3D network structure, good biocompatibility and high water content

Inactive Publication Date: 2010-10-20
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although photocrosslinked hydrogels have a short gel time (thus allowing rapid gel formation at the injected site by injection) and high mechanical strength (depending on the crosslink density), the photoinitiator necessary for the photocrosslinking reaction It is often cytotoxic, and the irradiation of ultraviolet light may also cause cell death at the irradiated site (Biomaterials, 2005, 26, 1211-1218; Biomaterials, 2009, 30, 344-353), which largely limits the Application of linked hydrogels as injectable in situ formable hydrogels

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1-3

[0024] Embodiment 1-3 provides the preparation method of silk fibroin (containing Tyr) hydrogel:

Embodiment 1

[0026] Put 20g silkworm raw silk into 4L Na 2 CO 3 (0.5wt%) degumming at 100° C. for 40 minutes in an aqueous solution, rinsed with deionized water after taking out, and dried in the air. Add 10 g of degummed silk solution to 100 mL of 9.3 mol / L LiBr aqueous solution at 40°C and keep stirring until the silk dissolves. Filter to remove undissolved silk. The obtained silk fibroin salt solution was dialyzed against deionized water to remove LiBr salt. The dialyzed silk fibroin solution is centrifuged to remove other insoluble impurities, and the supernatant is left, which is further concentrated to obtain a regenerated silk fibroin aqueous solution with a concentration of 12%.

[0027] Dissolve 5mg of horseradish peroxidase (HRP, 100unit / mg) in 2mL of 0.1mol / L PBS buffer solution (pH=6.0) to prepare a 2.5mg / mL HRP solution, and store it in a refrigerator at 4°C. Add 0.3 mL of HRP solution into 2.5 mL of regenerated silk fibroin aqueous solution (12%), mix well, and place in a...

Embodiment 2

[0029] Put 20g silkworm raw silk into 4L NaHCO 3 (0.5wt%) degumming at 100° C. for 40 minutes in an aqueous solution, rinsed with deionized water after taking out, and dried in the air. Add 10 g of degummed silk solution to 100 mL of 9.3 mol / L LiBr aqueous solution at 40°C and keep stirring until the silk dissolves. Filter to remove undissolved silk. The obtained silk fibroin salt solution was dialyzed against deionized water to remove LiBr salt. The dialyzed silk fibroin solution is centrifuged to remove other insoluble impurities, leaving the supernatant. The supernatant liquid is freeze-dried to obtain powder or foamy regenerated silk fibroin solid. Dissolve the regenerated silk fibroin solid in deionized water, centrifuge to remove insoluble matter, leave a clear liquid with a concentration of 15-20%, and dilute to the desired concentration (12% in this example) to obtain a regenerated silk fibroin aqueous solution.

[0030] Dissolve 0.5mg of horseradish peroxidase (HR...

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Abstract

The invention belongs to the technical fields of biotechnology, biological medical materials and tissue engineering, in particular to injected chemical crosslinking protein / polypeptide hydrogel and a preparation method thereof. The injected chemical crosslinking protein / polypeptide hydrogel consists of natural or synthesized protein / polypeptide with a crosslinking phenolic hydroxyl group, horse radish peroxidase and hydrogen peroxide which quickly form the chemical crosslinking hydrogel under physiological condition. The protein / polypeptide hydrogel has good biocompatibility and easily regulated mechanical strength, and is degradable, and the degraded product is amino acids capable of being absorbed by a human body. In addition, the framework of the protein / polypeptide hydrogel very approaches a protein three-dimensional network which supports the cell growth in extracellular matrix, and the invention can be widely applied to multiple biological and medical fields such as drug controlled-release carriers, cell culture supports and the like.

Description

technical field [0001] The invention belongs to the technical fields of biotechnology, biomedical materials and tissue engineering, and specifically relates to an injectable chemically cross-linked protein / polypeptide hydrogel and a preparation method thereof. Background technique [0002] In recent years, the development of new biomedical materials and related technologies has made significant progress in the diagnosis and treatment of many diseases. It is estimated that there are currently more than 8,000 medical devices, 2,500 diagnostic products, and 40,000 pharmaceutical preparations using various types of biomedical materials (including polymers, ceramics, and metals, etc.) (Adv. Mater., 2006, 18, 1345-1360). However, there is still an increasing demand for biomedical materials and their preparation technologies with better performance and more suitable for use in a biologically friendly environment. [0003] Hydrogel is a cross-linked network material rich in water,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08L89/00C08K3/20C08J3/24C08J3/075A61L27/22A61K47/42
Inventor 杨宇红龚祖光
Owner FUDAN UNIV
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