Photo-crosslinked recombinant collagen hydrogel, preparation method and application of the photo-crosslinked recombinant collagen hydrogel in 3D biological printing

A technology for recombinant collagen and bioprinting, which is used in applications, tissue regeneration, household appliances, etc., can solve the problems of gelatin having no rod-shaped triple-helix space structure, unfavorable industrialization promotion, complex processing conditions, etc., and achieves good formability and good formability. Good biological activity and physical and chemical properties

Inactive Publication Date: 2020-02-14
NANJING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] A large category of existing 3D bioprinting inks uses gelatin as the main raw material, but because of the breakage of covalent bonds and hydrogen bonds in the molecule, gelatin has no rod-like triple helix spatial structure, leaving only a quasi-triple helix structure, which depends on molecular Internal hydrogen bonds are maintained, making it lose its biological activity
At the same time, gelatin is insoluble in water, but can be dissolved by heating. It is a kind of heat-tolerant protein mixture. These make gelatin-based materials have high viscosity, complex processing conditions, high energy consumption, and unfavorable industrialization promotion (Ji Congling. Study on physical and chemical properties of silver carp skin gelatin and its performance improvement [D]. 2016. Master's degree thesis of Hefei University of Technology; Lin Shurong, Chen Bei, Wang Qin, et al. Research progress on collagen properties and preparation methods [J]. Fishery Research ,2018,40(05):73-81, etc.)

Method used

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  • Photo-crosslinked recombinant collagen hydrogel, preparation method and application of the photo-crosslinked recombinant collagen hydrogel in 3D biological printing
  • Photo-crosslinked recombinant collagen hydrogel, preparation method and application of the photo-crosslinked recombinant collagen hydrogel in 3D biological printing
  • Photo-crosslinked recombinant collagen hydrogel, preparation method and application of the photo-crosslinked recombinant collagen hydrogel in 3D biological printing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Preparation of photocrosslinked recombinant collagen hydrogel

[0043] Add 1 g of recombinant human collagen into 10 ml of phosphate buffer, stir for 30 minutes to dissolve it completely, and prepare a 10% (W / V) collagen phosphate solution. 0.6 ml of methacrylic anhydride was added to the collagen phosphate solution within 2 minutes, and stirred and reacted at 50° C. for 60 minutes. Add the reacted solution into a 10ml centrifuge tube, and centrifuge at 4000g for 3 minutes. Then take the supernatant and add deionized water to dilute it 2 times, adjust the pH value to 7, then add the diluted solution into a 36MM dialysis bag (MW: 14000), and dialyze under the condition of deionized water at room temperature for 7 days, every 8 hours Replace the deionized water. After the dialysis, the solution was frozen at -80°C, and then placed in a vacuum freeze dryer (Beijing Sihuan Scientific Instrument Factory Co., Ltd., model: LGJ-10D, freeze-dried according to the con...

Embodiment 2

[0047] Example 2 Preparation of photocrosslinked recombinant collagen hydrogel

[0048] Add 1 g of recombinant human collagen into 10 ml of phosphate buffer, stir for 30 minutes to dissolve it completely, and prepare a 10% (W / V) collagen phosphate solution. 0.6 ml of methacrylic anhydride was added to the collagen phosphate solution within 2 minutes, and stirred and reacted at 50° C. for 60 minutes. Add the reacted solution into a 10ml centrifuge tube, and centrifuge at 4000g for 3 minutes. Then take the supernatant and add deionized water to dilute it 2 times, adjust the pH value to 7, then add the diluted solution into a 36MM dialysis bag (MW: 14000), and dialyze under the condition of deionized water at room temperature for 7 days, every 8 hours Replace the deionized water. After the dialysis, the solution was frozen at -80°C, and then placed in a vacuum freeze dryer to freeze-dry to form a freeze-dried product.

[0049] Dissolve photoinitiator 2959 in deionized water to...

Embodiment 3

[0052] Example 3 Preparation of cell-loaded photocrosslinked recombinant collagen hydrogel

[0053] Add 1g of recombinant human collagen into 10ml of phosphate buffer, stir for 30 minutes to dissolve it completely, and configure it into a 10% (W / V) collagen phosphate solution; add 0.6ml of methacrylic anhydride to Collagen phosphate solution, stirred and reacted at 50°C for 60 minutes; put the reacted solution into a 10ml centrifuge tube, centrifuged at 4000g for 3 minutes; then took the supernatant and diluted it with deionized water 2 times to adjust the pH value 7. Then add the diluted solution into a 36MM dialysis bag (MW: 14000), and dialyze under deionized water at room temperature for 7 days, and replace the deionized water every 8 hours; after the dialysis, the solution is frozen at -80°C. Then place it in a vacuum freeze dryer for freeze-drying to form a freeze-dried product.

[0054] The blue photoinitiator LAP was dissolved in deionized water, and configured as a 0...

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Abstract

The invention discloses photo-crosslinked recombinant collagen hydrogel, a preparation method and application of the photo-crosslinked recombinant collagen hydrogel in 3D biological printing. The preparation method comprises the following steps: adding methacrylic anhydride into a phosphate solution of recombinant collagen, fully stirring and reacting to obtain methacrylic anhydride modified recombinant collagen, dissolving the modified recombinant collagen into a culture medium, adding a photoinitiator, uniformly mixing, then carrying out photoreaction, crosslinking and curing to obtain the photo-crosslinked recombinant collagen hydrogel. A recombinant collagen hydrogel precursor solution is placed in a mold or serves as 3D printing ink, and subjected to photoreaction, crosslinking and curing to prepare a tissue engineering scaffold or a 3D printing finished product; and cells can be added into the hydrogel precursor solution to prepare a tissue engineering scaffold or a 3D printing finished product loaded with the cells. The disclosed hydrogel is good in physical and chemical properties, good in biocompatibility, stable in quality and good in formability, can be prepared into anysize and shape, and is suitable for large-scale industrial production.

Description

technical field [0001] The invention belongs to the technical field of biomaterials, and relates to a photocrosslinked recombinant collagen hydrogel, a preparation method and its application in 3D bioprinting. Background technique [0002] Collagen is the most abundant structural protein in animals. Collagen has strong biological activity and biological function. It can participate in the migration, differentiation and proliferation of cells, so that connective tissue has mechanical strength. At the same time, collagen can also promote cell growth. It has good biocompatibility and biodegradable characteristic. Based on these characteristics, collagen is widely used in medical and health fields such as burn wound repair, corneal disease treatment, wound hemostasis, drug delivery and sustained release, orthopedics, health care, and beauty. Common collagen is isolated and extracted from animal tissues such as bovine bone, bovine hide, fish scale, fish skin, and pig skin. Ther...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J3/075C08J3/24C08L89/00C08H1/00A61L27/24A61L27/52A61L27/50A61K9/06A61K47/42C09D11/04B33Y70/00B33Y80/00B33Y10/00
CPCC08J3/075C08J3/24C08H1/00A61L27/24A61L27/52A61L27/507A61K9/06A61K47/42C09D11/04B33Y70/00B33Y80/00B33Y10/00C08J2389/00A61L2430/02A61L2430/16
Inventor 杨树林金明杰王子勋邓爱鹏朱帅
Owner NANJING UNIV OF SCI & TECH
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