Hydrogel material for promoting cartilage repair as well as preparation method and application of hydrogel material

A cartilage repair and hydrogel technology, applied in medical science, prosthesis, etc., can solve the problems of limited promotion and application potential, cumbersome experimental operation, and technical difficulty, so as to achieve in vivo repair, improve quality, and avoid ethical problems. Effects of related questions

Active Publication Date: 2022-04-12
孙烨
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

The disadvantage of this method is that it is necessary to construct a suitable DNA vector for encoding TGF-β protein, the experimental operation is cumbersome, the technology is difficult, and the potential for popularization and application is relatively limited; in addition, because the system contains DNA components, it may lead to genetic changes among different individuals. Risks posed by information transfer

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  • Hydrogel material for promoting cartilage repair as well as preparation method and application of hydrogel material
  • Hydrogel material for promoting cartilage repair as well as preparation method and application of hydrogel material
  • Hydrogel material for promoting cartilage repair as well as preparation method and application of hydrogel material

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preparation example Construction

[0033] In a first aspect, the present invention provides a method for preparing a cartilage-promoting hydrogel material, comprising the following steps:

[0034] (1) BMSCs are pretreated and cultured with transforming growth factor TGF-β, and the BMSCs are stimulated to secrete TGF-β-miRNA-extracellular vesicles after ingesting transforming growth factor TGF-β, and the TGF-β- miRNA-extracellular vesicles were filtered, centrifuged and concentrated, and the TGF-β-miRNA-extracellular vesicle suspension was collected;

[0035] (2) Co-cultivate TGF-β-miRNA-extracellular vesicle suspension and BMSCs to promote BMSCs to absorb TGF-β-containing miRNA-extracellular vesicles to obtain TGF-β-miRNA-extracellular vesicles soaked BMSC;

[0036] (3) Stir and mix the BMSCs containing TGF-β-miRNA-extracellular vesicles with the biocompatible hydrogel to obtain the cartilage-promoting hydrogel material.

[0037] In the present invention, the cartilage refers to a type of connective tissue, w...

Embodiment 1

[0055] Example 1 Preparation of Hydrogel Materials for Promoting Cartilage Repair

[0056] Step 1: Collect TGF-β3-miRNA-EV suspension

[0057] BMSCs were extracted from the femoral bone marrow cavity of healthy adult rats and placed in a medium containing 10% fetal bovine serum (FBS), 1% antibiotics (100U -1 mL Penicillin, 100mg -1 mL streptomycin and 0.25mg -1 mL amphotericin B) solution and 1×10 -3 mol sodium pyruvate (sodium pyruvate) in DMEM (Dulbecco's modified Eagle medium) medium; wherein, the environmental conditions of the cell incubator were set at a temperature of 37 ° C, CO 2 Concentration of 5% and humidity of 95%;

[0058] The DMEM medium was replaced every three days, and the cells were subcultured when the density of BMSC reached 80%;

[0059] When the BMSC proliferates to the third generation, in the above DMEM medium, an additional concentration of 50ng -1 mL of TGF-β3 solution, continue to culture for seven days to obtain TGF-β3-miRNA-extracellular ves...

Embodiment 2

[0073] Example 2 Application of Hydrogel Materials for Promoting Cartilage Repair in Rat Knee Articular Cartilage

[0074] Take 2mL of the cartilage-repairing hydrogel material prepared in Example 1 (the cell number of T3-EV-BMSC is 5×10 8 ), injected with a syringe to the injured part of the articular cartilage in the rat knee joint cavity, once a week, and injected continuously for eight weeks. The repair of articular cartilage in the rat knee joint cavity after eight weeks after operation is shown in figure 1 . from figure 1 The results can be seen that the surface of the damaged cartilage in the control group was filled with a large amount of granulation tissue, while the damaged cartilage in the knee joints of rats injected with the hydrogel material for promoting cartilage repair (T3-EV-BMSC hydrogel group) The surface of the site is filled with new tissue, the surface is smooth and flat, and there is no obvious boundary with the surrounding tissue. Therefore, the hy...

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Abstract

The invention belongs to the technical field of cartilage repair, and discloses a hydrogel material for promoting cartilage repair as well as a preparation method and application thereof. The preparation method comprises the following steps: pretreating BMSC (bone mesenchymal stem cells) by adopting TGF-beta3 to obtain TGF-beta3-miRNA-extracellular vesicles, co-culturing the BMSC and the TGF-beta3-miRNA-extracellular vesicles to obtain the BMSC of the TGF-beta3-miRNA-extracellular vesicles, and uniformly mixing the obtained product with hydrogel with biocompatibility to obtain the hydrogel material for promoting cartilage repair. According to the material, due to the synergistic effect of TGF-beta3-miRNA-extracellular vesicles, the limitation of promoting cartilage differentiation of BMSC by only using TGF-beta3 is overcome, the cartilage differentiation efficiency of BMSC is improved, and the material has a remarkable and excellent cartilage repair effect and can be applied to treatment of damaged cartilage.

Description

technical field [0001] The invention belongs to the technical field of cartilage repair, and in particular relates to a hydrogel material for promoting cartilage repair and its preparation method and application. Background technique [0002] Osteoarthritis (OA) is a degenerative joint disease characterized by the destruction of cartilage on the articular surface. OA is more common in middle-aged and elderly people and can seriously affect the quality of life. It is one of the main causes of disability in middle-aged and elderly people. At this stage, technical means aimed at repairing articular cartilage have become a new research direction for the prevention and treatment of OA. Articular cartilage has limited regeneration function due to low cell activity and no blood vessels to supply nutrients. Once damaged, it is difficult to reverse, which is also an important factor leading to the progression of OA. [0003] Currently, one way to repair damaged articular cartilage ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/12A61L27/22A61L27/38A61L27/50A61L27/52A61L27/54A61L27/58
Inventor 孙烨吴强尤永卿戴尅戎
Owner 孙烨
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