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Controllable self-crosslinking thiolated hyaluronic acid-collagen composite hydrogel and its preparation method and application

A composite hydrogel and hyaluronic acid technology, which can be used in pharmaceutical formulations, prostheses, drug delivery, etc., can solve the problems of easy-to-shrink cell adhesion, etc., and achieve good biodegradable properties, improved mechanical properties, and appropriate proportions Effect

Active Publication Date: 2020-11-10
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to overcome the deficiencies of the prior art, to provide a controllable self-crosslinking thiolated hyaluronic acid-collagen composite hydrogel and its preparation method and application, so as to effectively alleviate the existing gel materials used for cartilage repair. The existing deficiencies of easy shrinkage and poor cell adhesion

Method used

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  • Controllable self-crosslinking thiolated hyaluronic acid-collagen composite hydrogel and its preparation method and application
  • Controllable self-crosslinking thiolated hyaluronic acid-collagen composite hydrogel and its preparation method and application
  • Controllable self-crosslinking thiolated hyaluronic acid-collagen composite hydrogel and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] In this example, the preparation of mercaptolated hyaluronic acid, the steps are as follows:

[0047] (1) Dissolve sodium hyaluronate with a molecular weight of 0.3MDa in deionized water, add N-hydroxysuccinimide (NHS), fully dissolve, and then add 1-ethyl-(3-dimethylaminopropyl ) carbodiimide hydrochloride (EDC HCl), fully dissolved, adjust the pH value of the mixture to 4.75 with 1mol / L NaOH solution and 1mol / L HCl solution, react at room temperature for 2h, and then add cysteine Ammonia hydrochloride (CSH·HCl) solution was reacted at room temperature for 24 h, and then the pH value of the reaction solution was adjusted to 8.5 with 1 mol / L NaOH solution, and dithiothreitol (DTT) solution was added to react at room temperature for 12 h.

[0048] In this step, sodium hyaluronate, N-hydroxysuccinimide (NHS), 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC·HCl), semi The molar ratio of cystine hydrochloride (CSH·HCl) to dithiothreitol (DTT) is 1:2:4:4:12....

Embodiment 2

[0052] In this example, the controllable self-crosslinking mercaptolated hyaluronic acid-collagen composite hydrogel was prepared, and the steps were as follows:

[0053] (1) The HA-SH prepared in Example 1 is sterilized and dissolved in α-MEM medium to form a 10wt.% HA-SH solution; the α-MEM medium is α-MEM basal medium The mixture of penicillin-streptomycin (double antibody) and ascorbic acid was added on the basis of α-MEM medium. The volume concentration of double antibody in α-MEM medium was 1%, and the concentration of ascorbic acid was 50 μg / mL.

[0054] (2) Dissolve type I collagen with 0.5mol / L acetic acid solution under ice bath, adjust the pH value to 7.4 with 5mol / L NaOH solution, and then add PBS buffer solution with a concentration of 0.01mol / L and a pH value of 7.4 Adjust the concentration of type I collagen to 10 mg / mL to obtain a collagen solution; the type I collagen is extracted from bovine tendons.

[0055] (3) Draw the collagen solution and HA-SH solution...

Embodiment 3

[0065] In this example, the controllable self-crosslinking mercaptolated hyaluronic acid-collagen composite hydrogel was prepared, and the steps were as follows:

[0066] (1) According to the method in Example 1, the molar ratio of EDCI and CSA·HCl and the molecular weight of sodium hyaluronate are adjusted to prepare HA-SH with a grafting ratio of cysteine ​​of 33.54%, and the HA-SH Sterilize and dissolve in α-MEM medium to form a 1wt.% HA-SH solution; the α-MEM medium is based on the α-MEM basal medium by adding penicillin-streptomycin mixed solution ( double antibody) and ascorbic acid, the volume concentration of double antibody in α-MEM medium was 1%, and the concentration of ascorbic acid was 50 μg / mL.

[0067] (2) Dissolve type I collagen with 0.25mol / L acetic acid solution under ice bath, adjust the pH value to 7.8 with 5mol / L NaOH solution, and then add PBS buffer solution with a concentration of 0.01mol / L and a pH value of 7.8 Adjust the concentration of type I coll...

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Abstract

The invention provides controllable self-crosslinking sulfhydrylated hyaluronic acid-collagen composite hydrogel as well as a preparation method therefore. The composite hydrogel is prepared from hyaluronic acid hydrogel and I-type collagen or II-type collagen; the I-type collagen or II-type collagen is distributed in a three-dimensional crosslinking network structure. The composite hydrogel has rigidity and elasticity, is high in mechanical property, can promote spreading growth of cartilage cells, is favorable for phenotype maintaining for cells, can relieve the shortcomings that an existinggel material for cartilage reparation is easy to shrink and low in cell adhesion, and has an important application value in the field of cartilage injury reparation.

Description

technical field [0001] The invention belongs to the field of biological materials, and relates to a controllable self-crosslinking thiolated hyaluronic acid-collagen composite hydrogel and a preparation method thereof, and the controllable self-crosslinking thiolation hyaluronic acid-collagen composite hydrogel Applications in the field of bone repair. Background technique [0002] Articular cartilage defect is a common orthopedic disease. Since articular cartilage has no blood vessels, no lymph, and no nerves, it is difficult to repair itself. Existing treatment methods mainly rely on autologous chondrocyte transplantation, but are limited by the shortage of donor cells and easy differentiation during culture. As a cutting-edge technology, tissue engineering has attracted people's attention. The most basic principle of tissue engineering is that the synergistic growth factors of chondrocytes isolated and expanded in vitro are planted on the scaffold material, and after tr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08J3/075C08J3/24C08L5/08C08L89/00A61L27/48A61L27/38A61L27/58A61L27/50A61L27/52
CPCA61L27/3817A61L27/3852A61L27/48A61L27/50A61L27/52A61L27/58A61L2400/06A61L2430/06C08J3/075C08J3/24C08J2305/08C08J2389/00C08J2405/08C08J2489/00C08L5/08C08L89/00
Inventor 孙勇樊渝江陈亚芳贺萌萌张兴栋
Owner SICHUAN UNIV
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