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Collagen-based bio-ink for 3D bio-printing as well as preparation method and application of collagen-based bio-ink

A bio-ink and bio-printing technology, applied in biochemical equipment and methods, microorganisms, 3D culture, etc., can solve the problems of poor mechanical properties of pure collagen hydrogels, long collagen gelation time, and low structural resolution, etc. Achieve no cytotoxicity, prevent and treat cardiovascular diseases, and have good biocompatibility

Active Publication Date: 2021-07-23
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The main challenge currently facing 3D bioprinting is to prepare suitable bioinks
[0006] However, the collagen gelation time driven by temperature and cross-linking agent is longer, and the resolution of the structure after 3D printing is low; the mechanical properties of pure collagen hydrogel are poor, and it is difficult to form a 3D structure in a layer-by-layer manner (Osidak E O, Kozhukhov V I, Osidak M S, et al. Collagen as Bioink for Bioprinting: A Comprehensive Review [J]. International Journal of Bioprinting, 2020, 6(3)); this limits its application in the field of 3D bioprinting

Method used

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  • Collagen-based bio-ink for 3D bio-printing as well as preparation method and application of collagen-based bio-ink
  • Collagen-based bio-ink for 3D bio-printing as well as preparation method and application of collagen-based bio-ink
  • Collagen-based bio-ink for 3D bio-printing as well as preparation method and application of collagen-based bio-ink

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Embodiment 1: Preparation of collagen-based bioink

[0049] Dissolve 1 g of collagen in 400 mL of 10 mM hydrochloric acid solution, stir at 4°C for 8 h to dissolve completely, and prepare dilute hydrochloric acid solution of collagen. Use a molar concentration of 1M sodium hydroxide solution to adjust the pH of the collagen dilute hydrochloric acid solution to 9, add 2.3mL of methacrylic anhydride dropwise to the collagen dilute hydrochloric acid solution while stirring, and simultaneously add a molar concentration of 1M sodium hydroxide solution , the pH was stabilized at 9, and then stirred at 4°C and reacted in the dark for 8h. The reacted solution was poured into a dialysis bag with a cut-off flow rate of 8000-14000 Da, and dialyzed in 10 mM hydrochloric acid at 4° C. for 7 days, and the dialysate was changed twice a day. After the dialysis, the solution was frozen at -20°C, and then placed in a vacuum freeze dryer to freeze-dry to obtain a methacrylylated collagen...

Embodiment 2

[0053] Embodiment 2: the preparation of collagen-based bioink

[0054] Dissolve 1 g of collagen in 400 mL of 10 mM hydrochloric acid solution, stir at 4°C for 8 h to dissolve completely, and prepare dilute hydrochloric acid solution of collagen. Use a molar concentration of 1M sodium hydroxide solution to adjust the pH of the collagen dilute hydrochloric acid solution to 9, add 2.3mL of methacrylic anhydride dropwise to the collagen dilute hydrochloric acid solution while stirring, and simultaneously add a molar concentration of 1M sodium hydroxide solution , the pH was stabilized at 9, and then stirred at 4°C and reacted in the dark for 8h. The reacted solution was poured into a dialysis bag with a cut-off flow rate of 8000-14000 Da, and dialyzed in 10 mM hydrochloric acid at 4° C. for 7 days, and the dialysate was changed twice a day. After the dialysis, the solution was frozen at -20°C, and then placed in a vacuum freeze dryer to freeze-dry to obtain a methacrylylated coll...

Embodiment 3

[0057] Embodiment 3: the preparation of collagen-based bioink

[0058] The preparation method of the methacrylylated collagen freeze-dried sponge is the same as that in Example 2.

[0059] Dissolve 100 mg of methacrylylated collagen freeze-dried sponge in 5 mL of 0.2% acetic acid solution. After fully dissolving, adjust the pH to 7 with molar concentration of 1M sodium hydroxide, add photoinitiator LAP aqueous solution and proanthocyanidin aqueous solution to the solution, The concentration of LAP in the solution is 0.5% (w / v), and the concentration of proanthocyanidin is 0.008% (w / v). After mixing evenly, centrifuge to remove air bubbles to obtain collagen-based bioink, and place it in a refrigerator at 4°C for 24 hours.

[0060] Take an appropriate amount of collagen-based bio-ink in the mold, irradiate it with ultraviolet light with a power of 10W and a wavelength of 365nm, and it can be cured for 30-50s to form a hydrogel.

[0061] Collagen-based bioink of example 1-3 is...

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Abstract

The invention belongs to the technical field of biological materials, and particularly relates to collagen-based bio-ink for 3D biological printing as well as a preparation method and application of the collagen-based bio-ink. The specific preparation method of the collagen-based bio-ink comprises the following steps: adding methacrylic anhydride into a diluted hydrochloric acid solution of collagen for full reaction to obtain methylacryloylated collagen; and sequentially adding the methylacryloylated collagen into acetic acid, an ultraviolet light initiator LAP aqueous solution and a procyanidine aqueous solution, fully mixing, and cross-linking to obtain the collagen-based bio-ink. The collagen-based bio-ink prepared by the preparation method can be rapidly gelated within 30-50 seconds under ultraviolet irradiation, the formed hydrogel has good mechanical properties and good biocompatibility, and the collagen-based bio-ink can be used for 3D bio-printing.

Description

technical field [0001] The invention belongs to the technical field of biomaterials, and in particular relates to a collagen-based bioink for 3D bioprinting, a preparation method and application thereof. Background technique [0002] 3D bioprinting technology is driven by three-dimensional modeling programs such as CAD and CAM, and uses cells, proteins and biocompatible materials as raw materials to form three-dimensional entities in a layer-by-layer manner. In recent years, 3D bioprinting technology has developed rapidly. The emergence of 3D bioprinting technology has opened up new avenues for in vitro biomedical engineering, which can be used to construct tissue regeneration, in vitro biological models, and cell diagnosis. [0003] Bioink, the raw material for 3D bioprinting, is a cellular preparation suitable for processing by automated biomanufacturing techniques. The main challenge currently facing 3D bioprinting is the preparation of suitable bioinks. Bio-inks for 3...

Claims

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Application Information

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IPC IPC(8): C08L89/00C08K5/13C08H1/00C08J3/28C08J3/24C08J3/075B33Y70/10C12N5/00
CPCC08K5/13C08H1/00C08J3/28C08J3/24C08J3/075B33Y70/10C12N5/0062C08J2389/00C12N2533/54C12N2533/30C12N2537/10C08L89/00
Inventor 李立华吴姿琳李欣阳
Owner JINAN UNIVERSITY
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