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Collagen bio-ink for 3D printing and 3D printing method

A technology of collagen and bio-ink, which is applied in applications, inks, household appliances, etc., can solve the problems of poor safety, lack, and poor mechanical strength of medical materials, and achieve high safety, simple and gentle preparation methods, and good mechanical strength. Effect

Active Publication Date: 2021-07-06
胶原蛋白(武汉)生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] At present, there have been some attempts to use collagen as raw material for tissue / organ additive manufacturing, but there are still many problems: ①The preparation of most bio-inks is obtained by modifying methacrylic anhydride on the basis of collagen denaturation , leading to the lack of biological activity of the obtained bioink and the biomaterial prepared by 3D printing, and the inability to realize the functions of cell adhesion, migration, proliferation, etc.
For example, Chinese patent CN109337436A has prepared 3D printed bio-ink with methacrylated gelatin as raw material. Although this bio-ink can be used to prepare medical materials with good mechanical strength, this method uses denatured collagen (gelatin) as raw material, which loses The original high bioactivity of collagen cannot be used to print medical tissues / organs with excellent biological functions; similarly, Chinese patent CN110330797A prepared medical materials with better mechanical strength through photocrosslinking, which also uses methacrylation to modify Non-denatured gelatin, instead of using undenatured collagen with a complete triple helix structure, cannot be used to print medical tissues / organs with good biological functions
②When undenatured collagen is used as bioink, the mechanical strength of the printed medical materials is poor, and it is impossible to accurately print out medical tissues / organs with stable structures
Although this method maintains the biological activity of collagen, the mechanical strength of the prepared hydrogel is poor, and it is still impossible to realize the additive manufacturing of biological tissues / organs
③In order to increase the mechanical strength of medical materials, double crosslinking of chemical crosslinking agents is often required in the printing method, which will lead to the risk of residual toxicity of chemical crosslinking reagents, poor safety, and complicated preparation processes, such as , the literature (Materials Science and Engineering: C, 2020, 107: 110290) reported a double crosslinking method that continued chemical crosslinking after photocrosslinking
In summary, although collagen has great application prospects in 3D printing medical tissue or organ scaffolds, there is still a lack of collagen bioinks suitable for photocurable 3D printing. 3D printed bio-ink is a technical problem that urgently needs to be solved in this field

Method used

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  • Collagen bio-ink for 3D printing and 3D printing method
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  • Collagen bio-ink for 3D printing and 3D printing method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] The preparation of embodiment 1 collagen stock solution

[0042] Wash the yak tendon tissue to remove foreign matter and crush it into small pieces; degrease the crushed small pieces with 10% n-butanol, take the precipitate, wash it with water until neutral; decalcify the precipitate with 0.5M hydrochloric acid solution, After decalcification is completed, take the precipitate and wash it with water until neutral; soak the tissue precipitate in 0.1M sodium hydroxide solution, stir at 25°C for 4 hours, take the precipitate, wash it with water until neutral; use 0.5M pepsin containing 1g / L Collagen was extracted with acetic acid solution to obtain a crude collagen extract; pH was adjusted to neutral for enzyme inactivation; 8-14kDa dialysis bags were dialyzed to obtain collagen stock solutions with a concentration of 6 mg / mL.

Embodiment 2

[0043] 3D printing method one of embodiment 2 collagen bioink

[0044] 2.1 Collagen modified with methacrylic anhydride

[0045] Take the collagen stock solution prepared in Example 1, dilute it to a concentration of 1.5 mg / mL, add 150 mM NaCl and MOPS mixed buffer solution, adjust the pH to 7, and add 1 mL of methacrylic anhydride to every 200 mL of collagen stock solution under nitrogen protection , reacted in an ice bath to obtain a reaction product;

[0046] The reaction product obtained above was dialyzed with a mixed solution of 15 mM HCl and NaCl, and the dialysate was replaced for 6 times, followed by 15 mM HCl solution, and the dialysate was replaced for 5 times to obtain methacrylic anhydride-modified collagen ColMA , placed at -20°C for later use.

[0047] 2.2 Preparation of collagen bioink

[0048] Before use, take the above-mentioned ColMA solution, add photoinitiator LAP solution, until the final concentration of ColMA solution is 1.5 mg / mL, and the final conc...

Embodiment 3

[0050] Add the above configured collagen bio-ink into the light-curing 3D printer, and irradiate it with blue light with a wavelength of 405nm for 12s to perform light-curing 3D printing; after printing, use a blue-light flashlight with a wavelength of 405nm as the light source, with a distance of 5cm and light for 20s , and perform secondary curing to obtain 3D printed medical prosthetic ears. The printed medical prosthetic ears are as follows: figure 1 As shown in a, the prepared collagen bioink has fast curing speed and strong mechanical strength, and can print medical prosthetic ears similar to human ears. 3D printing method two of embodiment 3 collagen bioink

[0051] 3.1 Collagen modified with methacrylic anhydride

[0052] Take the collagen stock solution prepared in Example 1, dilute it to a concentration of 3 mg / mL, add 200 mM NaCl and MOPS mixed buffer solution, adjust the pH to 7, and add 1 mL of methacrylic anhydride to every 200 mL of collagen stock solution unde...

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Abstract

The invention belongs to the technical field of 3D printing bio-ink, particularly relates to collagen bio-ink for 3D printing, a preparation method of the collagen bio-ink and a method for 3D printing of biological tissues / organs by using the bio-ink, and provides bio-ink composed of ColMA and LAP and capable of being used for 3D printing of biological tissues / organs, wherein the bio-ink can be rapidly cured and formed under blue light irradiation and is excellent in printing performance. The invention further provides a simple and convenient 3D printing method of the collagen bio-ink. According to the invention, biological tissues / organs obtained by printing through the method have good mechanical strength, and the biological activity of collagen can be kept; and the collagen bio-ink capable of being used for photocuring 3D printing, provided by the invention, has a wide application prospect in the field of additive manufacturing for preparing biological tissues / organs with excellent biological activity and mechanical properties.

Description

[0001] This application claims the priority of the earlier application with the filing date of January 3, 2020, the application number of 2020100036085, and the title of the invention "a collagen bio-ink for 3D printing and a 3D printing method". The entire content of has been embodied in this application. technical field [0002] The invention belongs to the technical field of 3D printing bio-ink, and in particular relates to a collagen bio-ink for 3D printing and a method for 3D printing biomaterials. [0003] technical background [0004] 3D printing technology is an additive manufacturing technology based on precision and rapidity. By arbitrarily combining biological materials with chemicals or cells, etc., and printing layer by layer, a 3D model with precise structure and related biological functions can be obtained. 3D bioprinting is an interdisciplinary and interdisciplinary new regenerative medical engineering technology that reconstructs human tissues and organs by a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C09D11/04C08H1/00A61L27/24A61L27/50A61L27/54
CPCC09D11/04C08H1/00A61L27/24A61L27/50A61L27/54A61L2300/252A61L2430/14
Inventor 肖建喜杨霞霞
Owner 胶原蛋白(武汉)生物科技有限公司
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