In-situ cross-linking gel forming method of degradable protein

An in-situ cross-linking and protein technology, applied in the field of biomedical materials, can solve the problems of contamination, inability to perform simultaneous cross-linking of gels, unfavorable use, etc. sexual effect

Pending Publication Date: 2021-04-20
康膝生物医疗(深圳)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing ones are all cross-linked on dry protein materials, unable to perform synchronous in-situ cross-linked gels, and there will still be in the process of drying and transferring the dried materials to the container for light cross-linking. There is a risk of contamination, which is not conducive to later use

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Protein gel for injection

[0029] 1) Weigh an appropriate amount of pig type I collagen, disperse it in phosphate buffer (pH=7) and stir until the collagen is fully dissolved (dispersed) to prepare a 3% collagen solution (suspension), and then in Add hyaluronic acid to the solution and mix thoroughly, the final concentration of hyaluronic acid is 0.5%, marked as solution A;

[0030] 2) Add an appropriate amount of flavin adenine dinucleotide (FAD) to solution A, stir and mix thoroughly, the final concentration of FAD is 0.02%, marked as solution B;

[0031] 3) Fill solution B into a syringe and then inject it into the cartilage defect in the joint during arthroscopic surgery. Use the blue light beam (460nm) in the arthroscopic surgical instrument to irradiate the injection site with a radiation intensity of 50mW / cm 2 , the irradiation time is 10min.

Embodiment 2

[0032] Example 2: 3D printed gel scaffolds for tissue repair and regeneration

[0033] 1) Weigh an appropriate amount of bovine type I collagen, disperse it in phosphate buffer (pH 4.5-6.8) and stir well until the collagen is fully dissolved (dispersed) to prepare a 3% collagen solution (suspension), Then add hyaluronic acid and heparin into the solution, mix and stir thoroughly, the final concentration of hyaluronic acid is 1%, and the concentration of heparin is 0.2%, which is marked as solution A.

[0034] 2) Add the recombined growth factors bFGF and VEGF to the solution A and mix thoroughly, the final concentrations of bFGF and VEGF in the solution are 20 ng / ml and 10 ng / ml respectively. Then add an appropriate amount of riboflavin therein, and stir well so that the riboflavin is completely dissolved at a concentration of 0.1%, which is marked as solution B.

[0035] 3) Transfer solution B to the ink cartridge of the 3D bioprinter as the printed bioink.

[0036] 4) Impr...

Embodiment 3

[0037] Example 3: Tissue Repair and Regeneration 3D Printed Gel Scaffolds for Unloading Cells

[0038] 1) Weigh an appropriate amount of mouse type I collagen, disperse it in phosphate buffer (pH 4.0-6.5) and stir well until the collagen is fully dissolved (dispersed) to prepare a 2% collagen solution (suspension), Then add heparin into the solution and mix thoroughly, the concentration of heparin is 0.2%, marked as solution A.

[0039] 2) Add the recombined growth factors BMP-2 and VEGF to the solution A and mix thoroughly. The final concentrations of BMP-2 and VEGF in the solution are 20 ng / ml and 10 ng / ml respectively. Then add an appropriate amount of riboflavin therein, and stir well so that the riboflavin is completely dissolved at a concentration of 0.2%, which is marked as solution B.

[0040] 3) Extract human bone marrow and isolate bone marrow mesenchymal stem cells (BMSCs), culture them in vitro, and then collect BMSCs and disperse them into cell culture medium (1×...

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Abstract

The invention relates to an in-situ cross-linking gel forming method of degradable protein. The method comprises the following specific steps: S1, dispersing a degradable biological material taking protein as a main raw material in normal saline or a phosphate buffer solution to prepare a solution A; S2, adding riboflavin or riboflavin derivatives into the solution A, and stirring and mixing to prepare a solution B; S3, filling an injector or a 3D printer with the solution B; S4, performing in-vivo injection filling by using the injector, and irradiating the injected solution B by using an endoscope light source probe during injection; or adopting the 3D printer for printing, and adopting light beams emitted by a light beam generator for irradiating the solution B sprayed out of an ink nozzle of the 3D printer during printing. The gel is formed through in-situ cross-linking under the wet condition, the method is simple and easy to operate, no toxic cross-linking reagent is added, and the prepared gel can effectively improve the enzymolysis resistance of protein and has good biocompatibility.

Description

technical field [0001] The invention relates to the field of biomedical materials, in particular to an in-situ cross-linking gel forming method for degradable proteins. Background technique [0002] The degradation performance of biomaterials is directly related to the functionality and therapeutic effect of materials in vivo. In the fields of medical cosmetology and damaged tissue repair, in order to maintain the time of implanted materials in the body, materials are generally cross-linked. For example, in patent CN102924731B, in order to prolong the degradation time of collagen in vivo, aldehyde cross-linking agents, imine cross-linking agents and epoxy cross-linking agents are used to cross-link collagen. The use of these cross-linking agents can increase the degree of cross-linking of collagen and prolong the degradation time of collagen. However, these cross-linking agents remain in the material in the form of small molecules, which are difficult to remove and cause gr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J3/28C08J3/24C08J3/075A61L27/26A61L27/54A61L27/58A61K9/06A61K47/42A61K47/36C08L89/00C08L5/08
Inventor 邹杰刘瀚豪李鉴墨李罗浩
Owner 康膝生物医疗(深圳)有限公司
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