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Tissue engineered bone based on endochondral osteogenesis system and its construction method

A technology of endochondral ossification and tissue engineering bone, applied in prosthesis, artificially induced pluripotent cells, medical science, etc., can solve the problems of repair failure, ischemic necrosis of core parts, etc., and achieve good application prospects

Active Publication Date: 2015-12-02
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of this, the purpose of the present invention is to provide an endochondral osteogenesis system based on the problem that avascular necrosis often occurs in the core part of the large bone defect when the large tissue engineering bone is currently used, which easily leads to repair failure. Tissue engineered bone to avoid the problem of how to uniformly vascularize large tissue engineered bone and ensure sufficient nutrient supply

Method used

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  • Tissue engineered bone based on endochondral osteogenesis system and its construction method
  • Tissue engineered bone based on endochondral osteogenesis system and its construction method
  • Tissue engineered bone based on endochondral osteogenesis system and its construction method

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Example 1. Construction of tissue engineered bone based on endochondral osteogenesis system

[0023] 1. Culture of bone marrow MSCs

[0024] Use a heparin-treated syringe to extract 2-3 mL of human bone marrow under sterile conditions, immediately add it to a centrifuge tube filled with an equal amount of DMEM low-sugar medium, and quickly and gently blow it to make a cell suspension. Gently add to the centrifuge tube with an equal volume of lymphocyte separation medium, centrifuge at 2000r / min for 20min, collect the mononuclear cells in the buffy coat layer in another centrifuge tube, wash with DMEM (centrifuge at 1000r / min for 5min) twice, The supernatant was discarded, and the cells were resuspended in DMEM-L complete medium containing 15% fetal bovine serum, and diluted with 1×10 6 / mL cell density inoculated in a culture flask, recorded as the primary generation, placed at 37°C, 5% CO 2 Culture in a constant temperature incubator, change the medium every 3 days, ...

Embodiment 2

[0034] Example 2. In vivo ectopic osteogenesis of tissue engineered bone based on endochondral osteogenesis system

[0035] The tissue engineered bone (hypertrophy group and control group) based on the endochondral osteogenesis system constructed in Example 1 was implanted subcutaneously in nude mice, and after feeding for 8 weeks, the subcutaneous bone pieces were taken out and stained with SafraninO to observe the subcutaneous heterotopic bone in nude mice. The situation is formed, the result is as follows Figure 4 shown.

[0036] The results showed that after 8 weeks of in vivo culture in the hypertrophy group, bone tissue specimens were ectopically formed under the skin of the nude mice. Gross observation: wrapped in a film, translucent white, with a certain degree of toughness when touched. Histological observation: there are a small amount of fibrous cells and fibrous tissue wrapped around, lymphocytes can be seen but not immersed into the deep layer, there is a small...

Embodiment 3

[0037] Example 3. Repair of bone defect with tissue engineered bone based on endochondral osteogenesis system

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Abstract

The invention discloses a tissue-engineered bone based on an entochondrostosis system and a construction method thereof. The construction method is characterized in that mesenchymal stem cells are cultivated on a porous bone scaffold material to construct a tissue-engineering complex, subjected to in vitro cartilage differentiation induction cultivation for two weeks and continuously conducted with fat cartilage differentiation induction cultivation for two weeks to obtain the tissue-engineered bone. According to the invention, an in vivo research result shows that the in vivo ectopic bone formation can be realized, bone defects can be successfully repaired, the difficulties that the uniform vascularization of large piece of tissue-engineered bone cannot be realized and enough nutrition can't be supplied are solved, and the construction method has a very good application prospect in the construction of the tissue-engineered bone and bone defect repairing.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering, and relates to a tissue engineering bone and a construction method thereof. Background technique [0002] Bone defects caused by various traumas, infections, and tumor resections are common clinical diseases. At present, autologous bone, xenograft bone, allogeneic demineralized bone, or artificial bone substitutes are usually used to repair bone defects clinically. Autologous bone grafting has osteoconduction and osteoinductive effects, and there is no risk of disease transmission, but its application is limited due to problems such as limited sources, inability to cut and shape at will, morphological and functional defects of the remaining donor site, and the inability to predict the amount of bone resorption after surgery. . In order to solve these problems caused by autologous bone transplantation, a variety of bone substitute materials have been developed, such as xenograft bone, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/074A61L27/38A61L27/56
Inventor 董世武康菲窦策杨小超李建美曹震
Owner ARMY MEDICAL UNIV
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