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Method for preparing collagen-II aquagel for inducing chondrogenic differentiation of stem cells

A collagen hydrogel and cartilage-forming technology, which is applied in the field of biomedical materials, can solve the problems that cells cannot provide a suitable environment for growth and differentiation, the sponge structure lacks three-dimensional cartilage microenvironment bionics, and the important role of type II collagen is not enough to highlight the important role of collagen, etc. Achieve good cytocompatibility, promote chondrogenic differentiation, and non-toxic raw materials

Active Publication Date: 2018-05-08
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the scaffold material prepared by the above method has potential cytotoxicity due to the toxicity of glutaraldehyde, genipin, and EDC / NHS; and the sponge structure lacks bionics for the three-dimensional cartilage microenvironment, and cannot provide cells with a suitable environment. Growth and Differentiation Environment
Furthermore, the use of composite gel materials is insufficient to highlight the important role of type II collagen in influencing and directing cell behavior

Method used

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  • Method for preparing collagen-II aquagel for inducing chondrogenic differentiation of stem cells
  • Method for preparing collagen-II aquagel for inducing chondrogenic differentiation of stem cells
  • Method for preparing collagen-II aquagel for inducing chondrogenic differentiation of stem cells

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Embodiment 1

[0037] The steps of this embodiment are as follows:

[0038] (1) Preparation of type II collagen solution

[0039] Under the condition of normal pressure and 4°C, the type II collagen was prepared into a type II collagen solution with a concentration of 2 mg / mL with 20 mM hydrochloric acid solution, and stirred for 12 hours to dissolve the II collagen.

[0040] (2) Synthesis of photocrosslinkable type II collagen

[0041] At normal pressure at 4°C, use 0.2M disodium hydrogen phosphate buffer and 5M sodium hydroxide solution to adjust the pH value of the type II collagen solution to 7.5, and then adjust the molar ratio of amino groups to methacrylic anhydride in the collagen to be 1 : 5, add methacrylic anhydride to the type II collagen solution after the pH value adjustment, continue to stir and react for 4 hours, supplement 5M sodium hydroxide solution in the reaction process to keep the pH value of the reaction system at 7.5.

[0042] After the reaction is finished, remove...

Embodiment 2

[0047] The steps of this embodiment are as follows:

[0048] (1) Preparation of type II collagen solution

[0049] Under the condition of normal pressure and 0°C, the type II collagen was prepared into a type II collagen solution with a concentration of 3 mg / mL with 20 mM acetic acid solution, and stirred for 12 hours to dissolve the II collagen.

[0050] (2) Synthesis of photocrosslinkable type II collagen

[0051] Under normal pressure at 0°C, use 0.5M sodium dihydrogen phosphate buffer and 3M sodium hydroxide solution to adjust the pH value of the type II collagen solution to 8, and then adjust the molar ratio of amino groups to methacrylic anhydride in the collagen to be 1 : 2.5, add methacrylic anhydride to the type II collagen solution after the pH value adjustment, continue to stir and react for 8 hours, supplement 3M sodium hydroxide solution in the reaction process to keep the pH value of the reaction system at 8.

[0052] After the reaction is finished, remove unre...

Embodiment 3

[0057] The steps of this embodiment are as follows:

[0058] (1) Preparation of type II collagen solution

[0059] Under the condition of normal pressure and 4°C, the type II collagen was prepared into a type II collagen solution with a concentration of 4 mg / mL with 100 mM hydrochloric acid solution, and stirred for 12 hours to dissolve the II collagen.

[0060] (2) Synthesis of photocrosslinkable type II collagen

[0061] Under normal pressure at 4°C, adjust the pH value of the type II collagen solution to 7.5 with 1M disodium hydrogen phosphate buffer solution and 1M sodium hydroxide solution, and then according to the molar ratio of amino group and methacrylic anhydride in the collagen is 1: 0.5, add methacrylic anhydride to the pH-adjusted type II collagen solution, and continue to stir for 12 hours. During the reaction, add 1M sodium hydroxide solution to keep the pH of the reaction system at 7.5.

[0062]After the reaction is finished, remove unreacted methacrylic anhy...

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Abstract

The invention discloses a method for preparing collagen-II aquagel for inducing chondrogenic differentiation of stem cells. The method comprises the steps: firstly, synthesizing photo-crosslinking collagen II, which has photo-crosslinking performance and keeps tri-helical conformation of collagen II complete, through an amidation reaction between lateral-chain lysine epsilon-amino of the collagenII and methacrylic anhydride; and then, initiating the polymerization crosslinking of the photo-crosslinking collagen II under the action of a photoinitiator so as to form the collagen-II aquagel withcertain mechanical strength and adjustable mono-components. The collagen-II aquagel is used for providing a biological bionic differentiation microenvironment for the stem cells.

Description

technical field [0001] The invention belongs to the technical field of biomedical materials, and relates to a preparation method and application of an inductive hydrogel material used for cartilage defect repair. Background technique [0002] Given that articular cartilage tissue is avascular and has a low overall metabolic rate and viable cell density, its ability to self-repair is limited when patients suffer from cartilage-related diseases or trauma. Common clinical methods for cartilage repair include bone marrow stimulation, osteochondral transplantation and prosthetic replacement, etc., but these methods usually cause the formation of fibrocartilage tissue, because the structural composition and mechanical strength of fibrocartilage tissue are different from natural hyaline cartilage tissue. Therefore, it is difficult for common clinical methods of cartilage repair to achieve long-term effective therapeutic effects. Cell therapy based on tissue engineering, by inocula...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J3/28C08J3/075C08L89/00C08L51/00C08F289/00C08F220/08A61L27/24A61L27/50A61L27/52
CPCA61L27/24A61L27/50A61L27/52A61L2430/06C08F289/00C08J3/075C08J3/28C08J2389/00C08J2451/00C08F220/08
Inventor 孙静杨棵卫丹范红松张兴栋
Owner SICHUAN UNIV
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