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Photocuring enzyme cross-linking bioink, preparation method of bioink and 3D printing method

A bio-ink and 3D printing technology, applied in the field of bio-3D printing, can solve the problem of single cross-linking method, 3D bionic structure is difficult to take into account cell compatibility, mechanical strength and degradability, single component of bio-3D printing ink, etc. problem, to achieve the effect of high cytocompatibility, high degradability and high cell adhesion

Inactive Publication Date: 2020-02-04
SOUTHERN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] One of the objectives of the present invention is to provide a light-curing enzyme cross-linked bio-ink and its preparation method to solve the problem that the bio-3D printing ink has a single component and a

Method used

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  • Photocuring enzyme cross-linking bioink, preparation method of bioink and 3D printing method
  • Photocuring enzyme cross-linking bioink, preparation method of bioink and 3D printing method
  • Photocuring enzyme cross-linking bioink, preparation method of bioink and 3D printing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] A preparation method of photocurable enzyme cross-linked bio-ink, comprising the steps of:

[0037] S1, preparing fibrin stock solution and methacrylic anhydride gelatin solution in advance;

[0038] The preparation process of fibrin stock solution is to mix sterile fibrinogen powder with 1×~10× cell culture medium solution, shake and dissolve at 0℃~40℃ to obtain fibrin stock solution, and the obtained fibrin stock solution is in- Store at 80℃~37℃ for later use;

[0039] The preparation of methacrylic anhydride gelatin liquid is to blend methacrylic anhydride gelatin and photoinitiator powder, the weight ratio of photoinitiator to methacrylic anhydride gelatin is 0.01~50:1, and then add 1×~10 X’s cell culture medium solution was stirred at 0°C to 80°C and protected from light for 0.5 to 6 hours to obtain a methacrylic anhydride gelatin solution, and the obtained methacrylic anhydride gelatin solution was kept at -80°C to 37°C in the dark. Store at ℃ for later use;

...

Embodiment 2

[0046] A method for 3D printing of a biomimetic structure using photocurable enzyme cross-linked bio-ink, comprising the steps of:

[0047] (1) Load the photocurable enzyme cross-linked bio-ink into the printing silo of the 3D printer, set the temperature of the silo to 0°C to 45°C, the moving speed of the nozzle to 0-100mm / s, and the extrusion pressure to 0-100mm / s 25MPa, printing platform temperature is 0°C to 45°C;

[0048] (2) Print according to the required digital model, irradiate with ultraviolet light while printing, and perform photocrosslinking on the printed structure; the wavelength of ultraviolet light used is 500-315nm, and the light intensity is 5-500 mw / cm 2 , the irradiation time is 1-90s; (3) Add cross-linking solution on the surface of the printed structure, so that the cross-linked solution completely immerses the printed structure, and put it into the incubator for 10 minutes to 2 hours;

[0049] (4) Transfer the printed structure to a complete medium and...

Embodiment 3

[0055] A photocurable enzyme cross-linked bio-ink described in this example has the same composition and preparation method as in Example 1, the difference being that in the photo-cured enzyme cross-linked bio-ink, the mass percentage of fibrinogen 1-10 wt%, the mass percentage of methacrylic anhydride gelatin is 0.1-15 wt%, and the mass percentage of photoinitiator is 0.1-1 wt%.

[0056] It can be obtained from experiments that reducing the amount of solid materials in the bioink formulation does not affect the high cytocompatibility, high mechanical strength, high degradability and non-cytotoxic properties of the double network hydrogel system, in which bioactive cells Proliferation and differentiation were good.

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Abstract

The invention discloses photocuring enzyme cross-linking bioink, a preparation method of the bioink and a 3D (three dimensional) printing method. The photocuring enzyme cross-linking bioink comprisesthe following ingredients: 0.1-50wt% of fibrinogen, 0.1-50wt% of methacrylic anhydride modified gelatin and 0.01-20wt% of photoinitiator. The photocuring enzyme cross-linking bioink has diversified material components; enzyme cross-linking of the fibrinogen is added based on ultraviolet light cross-linking of methacrylic anhydride modified gelatin; and a double-network hydrogel system comprising bioactive cells is constructed. A bionic structure printed by the photocuring enzyme cross-linking bioink has the characteristics of high cell compatibility, high mechanical strength, high degradability and no cytotoxicity, provides a good bionic environment for proliferation, differentiation and protraction of the bioactive cells and is applicable to the field of tissue engineering such as biological tissue construction and repair.

Description

technical field [0001] The invention relates to the technical field of biological 3D printing, in particular to a photocurable enzyme cross-linked bio-ink, a preparation method thereof and a 3D printing method. Background technique [0002] As one of the preparation methods of biomimetic structures, 3D bioprinting technology has the advantages of precisely controlling the morphology and cell distribution of biomimetic structures. Bio-3D printing technology can precisely control the printing structure in three-dimensional space, and can print complex bionic structures according to the unique anatomical and physiological standards of the human body, providing a new technical means for the medical field of tissue engineering and clinical repair of tissue or organ defects. [0003] Existing bio-3D printing inks have a single component and usually only contain a hydrogel system. The cross-linking method is also relatively simple, and usually only one of chemical cross-linking, p...

Claims

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Application Information

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IPC IPC(8): A61L27/40A61L27/22A61L27/52A61L27/58B29C64/124B29C64/379B33Y10/00B33Y40/00B33Y70/00
CPCA61L27/222A61L27/227A61L27/52A61L27/58B33Y10/00B33Y40/00B33Y70/00B29C64/124B29C64/379C08L89/00
Inventor 黄文华吴耀彬李婷
Owner SOUTHERN MEDICAL UNIVERSITY
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