Preparation method of tissue engineering bone cartilage compound and its application

A tissue-engineered bone and composite technology, used in biochemical equipment and methods, bone implants, tissue cell/virus culture devices, etc., to achieve the effects of promoting early formation, delaying degeneration, and cartilage stabilization

Inactive Publication Date: 2007-05-02
INST OF HEMATOLOGY & BLOOD HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] The technical problem to be solved by the present invention is to provide a method of using pluripotent stem cells to construct excellent tissue engineered osteochondral...

Method used

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  • Preparation method of tissue engineering bone cartilage compound and its application
  • Preparation method of tissue engineering bone cartilage compound and its application
  • Preparation method of tissue engineering bone cartilage compound and its application

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preparation example Construction

[0038] As shown in Figure 1, the tissue engineering osteochondral compound preparation method of the present invention comprises the following steps:

[0039]1) Use the method of density gradient centrifugation of lymphocyte separation medium to isolate and expand bone marrow pluripotent stem cells BMSCs in vitro. When BMSCs reach confluence at passage 2-3, digest and count according to 3×10 7 -6×10 7 / 100mm 2 Density inoculation, using different cartilage and osteogenesis induction systems for initial induction of cartilage and bone origin;

[0040] 2) taking fresh ilium and knee joints of New Zealand rabbits, and performing decalcification treatment to prepare decalcified bone matrix;

[0041] 3) Then the demineralized bone matrix and type I collagen sponge were cut into cubes of required size, pre-coated with human fibronectin and dried overnight;

[0042] 4) BMSCs cultured in different induction systems in step 1 were respectively inoculated into the decalcified bone ma...

Embodiment 1

[0046] Example 1 Using mesenchymal stem cells to prepare tissue-engineered cartilage to repair articular cartilage defects

[0047] 1) In vitro isolation and expansion of bone marrow pluripotent stem cells: extract rabbit bone marrow through bone marrow puncture, and use density gradient centrifugation to separate mononuclear cells with lymphocyte separation medium (specific gravity: 1.077) 7 / 100mm 2 The density was inoculated in culture flasks, and the culture medium contained 60% DMEM-LG / F / 12, 40% MCDB-201 (Gibco), 2% FBS, 10ng / ml EGF, 10ng / ml PDGF-bb (Peprotech) components . Place at 37°C, 5% CO 2 cultured in an incubator. After 24 to 72 hours, discard the suspended cells, add fresh medium to continue culturing, and change the medium every 3 days until the cells grow to 80% confluence, digest the cells with 0.25% trypsin-0.1% EDTA at 37°C, Count to 3 x 10 4 -5×10 4 / cm 2 Cells were inoculated at a certain density, and cultured and subcultured according to the above ...

Embodiment 2

[0052] Example 2 Using Bone Marrow Mesenchymal Stem Cells to Prepare Tissue-Engineered Osteochondral Compounds to Repair Articular Cartilage Defects

[0053] 1) In vitro isolation and expansion of bone marrow pluripotent stem cells: the rabbit bone marrow was extracted by bone marrow puncture, and the bone marrow mononuclear cells were separated by lymphocyte separation medium. 7 / 100mm 2 The density was inoculated in culture flasks, and the culture medium contained 60% DMEM-LG / F / 12, 40% MCDB-201 (Gibco), 2% FBS, 10ng / ml EGF, 10ng / ml PDGF-bb (Peprotech) components . Place at 37°C, 5% CO 2 cultured in an incubator. After 2-3 days, discard the suspended cells, add fresh medium to continue culturing, and change the medium every 3 days until the cells grow to 80% confluence, digest the cells with 0.25% trypsin-0.1% EDTA at 37°C, and count , to 3 x 10 4 -5×10 4 / cm 2 Cells were inoculated at a certain density, and cultured and subcultured according to the above conditions. ...

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Abstract

A tissue-engineered bone-cartilage assembly for repairing the damaged bone and cartilage by inserting it in the damaged position is prepared through preparing bone and cartilage by tissue engineering, adhering them together, and making a wedge on bone part.

Description

technical field [0001] The invention relates to a preparation method and application of a tissue engineering compound, in particular to a preparation method and application of a tissue engineering osteochondral compound. Background technique [0002] The injury and defect of articular cartilage is a common disease in clinical orthopedics. Connection replacement and osteochondral defect reconstruction are very important for restoring bone and cartilage function. At present, non-biological substitutes, xenogeneic and allogenic osteochondral transplantation are usually used clinically. for the treatment of cartilage defects. However, the connection replacement effect of non-biological substitutes is not long-lasting, and often wears and loosens; the availability of autologous donor tissue and the damage of the donor limit the application of autologous transplantation, and there are risks such as immune rejection and viral infection in allogeneic transplantation , cannot achiev...

Claims

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Application Information

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IPC IPC(8): A61L27/00A61L27/38A61L27/40A61L31/00C12M3/00A61B17/56A61F2/28
Inventor 葛薇李长虹
Owner INST OF HEMATOLOGY & BLOOD HOSPITAL CHINESE ACAD OF MEDICAL SCI
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