Silk fibroin 3D printing bio-ink and application thereof

A 3D printing and silk fibroin technology, applied in medical science, prosthesis, additive processing, etc., can solve problems such as the dependence of microscopic pore size and porosity, affecting cell adhesion and growth, and difficult to process three-dimensional scaffolds. Achieve good biocompatibility and mechanical properties, improve material exchange efficiency, and achieve personalized regulation

Inactive Publication Date: 2021-06-18
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although 3D printing technology has solved the problem of pores in the macroscopic millimeter scale, the size and porosity of the pores in the microscopic scale still depend on the bioink.
For particle pore-making technology, scaffolds with high porosity and large pore diameter can be prepared, but the connectivity between the internal pores is poor, and the pore-forming particles are not easy; although extrusion molding technology can achieve personalized controllability, However, it is still difficult to process three-dimensional scaffolds with complex shapes
In addition, most of the related synthetic polymer bioscaffolds have the disadvantage of poor biocompatibility
Therefore, the above problems in the prior art affect the adhesion and growth of cells in tissue regeneration materials, as well as the application of biological scaffolds in tissue engineering and biomedicine, and urgently need to be solved.

Method used

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  • Silk fibroin 3D printing bio-ink and application thereof
  • Silk fibroin 3D printing bio-ink and application thereof
  • Silk fibroin 3D printing bio-ink and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] The preparation of embodiment 1 silk fibroin solution

[0065] (1) Silk degumming method: Measure 12L of pure water and pour it into the degumming pot, heat the pure water, weigh 25.44g of anhydrous Na 2 CO 3 , when the water is about to boil add. Weigh 30g raw silk, wait for anhydrous Na 2 CO 3 After fully dissolving in boiling water, add and cook for 30 minutes. After cooking, take it out, wash it with 2L of pure water repeatedly for 3 times, and put it in a fume hood to dry;

[0066] (2) Silk dissolving method: Weigh 80.75g lithium bromide (99.9%) in a 500mL beaker, set the volume to a 100mL volumetric flask (9.3M), weigh 5 parts of degummed silk in step (1), each 5g Put it in a 50mL beaker, pour 20mL lithium bromide solution into each beaker, place it in a 60°C oven, shake the beaker every 1 hour, and take it out after 4 hours;

[0067] (3) Silk fibroin solution dialysis method: cut out 5 sections of dialysis bag, and the molecular weight cut-off of the dialys...

Embodiment 2

[0070] Embodiment 2: Preparation of photosensitive silk fibroin

[0071] (1) Add 424mM GMA solution to the silk fibroin solution obtained in step (2) of Example 1, mix well, and react at 60° C. for 3 hours at a speed of 30 rpm to produce a high-yield reaction.

[0072] (2) After filtering, use a dialysis bag with a molecular weight cut-off of 12-14kDa to dialyze for 4 days, replace the deionized water 6 times a day, then place it in a -80°C refrigerator overnight, freeze-dry it and place it at -4°C for future use. use.

Embodiment 3

[0073] Example 3: Cell Culture

[0074] (1) Primary acquisition of BMSCs (Bone Marrow Mesenchymal Stem Cells)

[0075] Pour ether into cotton to anesthetize SD rats, kill them by necking, soak the dead bodies of rats in 75% ethanol for disinfection; soak them for 30 minutes, take them out and put them in the ultra-clean workbench to start the experiment, cut the skin of the legs of the rats and muscles, remove the complete tibia and femur, soak in PBS solution containing 10% double antibody for 5-10 minutes, take it out, soak it in PBS solution containing 5% double antibody for 5-10 minutes, and finally soak it in In PBS solution containing 1% double antibody for 5-10 minutes; cut off both ends of the tibia and femur to expose the bone marrow cavity, insert 1mL filled with complete medium into the end of the bone marrow cavity, and slowly push the syringe. Place the centrifuge tube under the other end of the bone to collect the flushed fluid. After all flushing is completed,...

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Abstract

The invention relates to silk fibroin 3D printing bio-ink and application thereof. The 3D printing bio-ink comprises a silk fibroin-based material, particles and an additive, and the additive is used for crosslinking and curing the silk fibroin 3D printing bio-ink. Based on the pore problem of an existing 3D printing scaffold material, a material is added into the bio-ink, so that the self-sustaining property of a 3D structure in the 3D printing process is improved, meanwhile, the material is removed after printing, holes of 20-500 microns are formed in the 3D printing structure, the diameter of 50% or more of the holes is larger than or equal to 100 microns, the exchange efficiency of substances in the scaffold is greatly improved, and the cell proliferation efficiency and differentiation capacity are improved. The method created by the invention and the 3D printing bionic structure prepared according to the method have very important significance on research on the action mechanism between materials and cells, tissue regeneration and biomedical materials.

Description

technical field [0001] The invention relates to 3D printing bio-ink, in particular to a silk fibroin 3D printing bio-ink and its application. Background technique [0002] Currently, tissue engineering is considered to be one of the most promising ways to completely solve the problem of tissue and organ repair. The core of tissue engineering is to construct a complex composed of cells and cell scaffolds. Among them, the tissue engineering scaffold is used as a place for planting cells and a template for tissue regeneration. The size and porosity of the internal pores have an important impact on the adhesion and growth of cells, and ultimately determine the repair effect of the site to be repaired. The combination of 3D printing and tissue engineering can achieve higher precision fabrication of complex tissues, realize the feasibility of spatial arrangement of cells, and control porosity to maintain nutrient exchange and cell survival. Although 3D printing technology solves...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/22A61L27/56B33Y10/00B33Y30/00B33Y70/10
CPCA61L27/227A61L27/56B33Y70/10B33Y30/00B33Y10/00C08L89/00
Inventor 郑兆柱陈莉汪涛吴佳阳胡涛关晋平赵伟李刚王晓沁
Owner SUZHOU UNIV
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