Tissue engineering scaffold material for repairing cartilage defects and preparation method thereof

A tissue engineering scaffold and soft tissue technology, applied in the preparation of the scaffold material, in the field of tissue engineering scaffold materials, can solve the influence of demineralized bone matrix on the ability to repair cartilage, the limited source of allogeneic human bone materials, and the inconsistency in repairing bone and cartilage. and other problems, to achieve the effect of low immunogenicity, good cell and biocompatibility, and good biocompatibility

Inactive Publication Date: 2011-02-23
昆明医学院第一附属医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the prior art, due to the different decalcification intensity and time of DBM, the effect of repairing bone and cartilage is inconsistent.
The inventors of the present application have found after research that an appropriate amount of decalcified bone matrix is ​​beneficial to cartilage formation, while excessive decalcified bone matrix reduces its ability to form cartilage. Part of the reason may be that an appropriate amount of decalcified bone matrix can make the bone contained in the bone matrix The exposure of bioactive factors such as morphogenetic protein (BMP) and TGF-β, while excessive decalcification affects the activity of demineralized bone matrix, thus affecting the ability of demineralized bone matrix to repair cartilage
In addition, at home and abroad, bovine bones, rabbit bones, and sheep bones are mainly used as raw materials at present, and the structure of these heterogeneous bone materials is far from that of humans, and the source of allogeneic human bone materials is limited, and it is easy to spread hepatitis, AIDS, etc.

Method used

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  • Tissue engineering scaffold material for repairing cartilage defects and preparation method thereof
  • Tissue engineering scaffold material for repairing cartilage defects and preparation method thereof
  • Tissue engineering scaffold material for repairing cartilage defects and preparation method thereof

Examples

Experimental program
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Effect test

experiment example 1

[0044] Experimental Example 1: Visual and SEM observation of DSBM

[0045] The appearance of DSBM scaffold material is white and sponge-like by naked eye. When it absorbs water, it has good elasticity when clamped with tweezers. It can squeeze out water and restore it. It has a certain toughness. It maintains its original shape and has a certain strength when dry. Porous structure, the surface pores are connected to the deep pores, and the pore sizes are different ( figure 1 ). Take 10 blocks of the prepared DSBM, fix with 2% glutaraldehyde at 4°C for 24 hours, rinse with 0.1 mol / L sodium cacodylate buffer, and fix with 1% osmium tetroxide for 2 hours. Rinse with 0.1mol / L sodium cacodylate buffer solution, dehydrate step by step with ethanol, store in isoamyl acetate overnight, dry at the critical point of carbon dioxide, adhere to the stage in a direction, spray gold in vacuum, and observe on the machine. The surface structure of DSBM was observed by scanning electron micro...

experiment example 2

[0048] Experimental example 2: In vitro co-cultivation of DSBM and BMSCs

[0049] According to the above method, 24 pieces of DSBM were taken from the three time periods of decalcification 6h, 12h and 24h respectively. After 24 hours of culture, the culture medium was sucked off with sterile absorbent cotton balls, and then placed in six 24-well culture plates. Put 1 piece of decalcified 6h DSBM material in holes A1, B1, C1, and D1 of each hole, put 1 piece of decalcified 12h DSBM material in holes A2, B2, C2, and D2, and put 1 piece of decalcified 12h DSBM material in holes A3, B3, C3, and D3. Calcium 24h DSBM material, will have a density of 5 x 10 6 40 μL of third-generation bone marrow mesenchymal stem cells (BMSCs) / mL were inoculated on the pre-wet material of 24-well culture plate, CO 2 Cultivate in the incubator for 4 hours, then add 1.2 mL of L-DMEM medium containing 15% newborn bovine serum to continue the culture, add the same medium as the background to wells A4, B...

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Abstract

The invention discloses a tissue engineering scaffold material for repairing cartilage defects and a preparation method thereof. The tissue engineering scaffold material is prepared by the following steps of: cleaning the spongy bone at the shoulder blade of a fresh pig; cutting the spongy bone into small blocks with diameter of between 5 and 10mm and thickness of between 3 and 5mm; degreasing, decalcifying and deproteinizing; immersing with alcohol for 30 minutes; and air-drying, wherein the decalcifying time is 6 hours. The prepared demineralized spongy bone matrix (DSBM) has the characteristics of relatively simple and convenient preparation method, and capacity of being obtained on a large scale, can be stored for a long time at a low temperature and is economic and cheap. The material can be prepared into different shapes and sizes according to the shapes of the defects to be repaired, so the material is convenient to apply. The DSBM remains a natural netlike gap structural system of the pig spongy bone, has high cellular compatibility, biocompatibility and degradability and is a very good cartilage tissue engineering scaffold material.

Description

technical field [0001] The invention belongs to the technical field of cartilage defect repair materials, and in particular relates to a tissue engineering support material for cartilage defect repair. At the same time, the invention also relates to a preparation method of the bracket material. Background technique [0002] Defects of articular cartilage caused by trauma and degeneration are very common in clinical practice. After articular cartilage is damaged, its self-repair ability is limited. How to repair the damage of articular cartilage and subchondral bone, promote the self-repair of articular cartilage, and rebuild joint function has become a problem for clinicians and experts. Important questions for researchers to address. [0003] Arthroplasty, subchondral drilling, or microfracture, etc., generate fibrocartilage repair by migrating various progenitor cells in the blood and bone marrow to the articular cartilage defect. Although fibrocartilage is very differen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/56A61L27/24A61L27/36
Inventor 李彦林刘流韩睿王福科金耀峰陈建明
Owner 昆明医学院第一附属医院
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