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Construction method of sclerotomal cell idiosyncratic transcription factor - 2 gene decorated tissue engineered bone

An osteoblast-specific, transcription factor technology, applied in the field of bone tissue engineering, can solve problems such as reducing antigenicity, reducing mechanical strength, and reducing material osteoinductivity

Inactive Publication Date: 2006-02-15
ARMY MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the current bio-derived materials also have their disadvantages: the deproteinization treatment better retains the pore structure and mechanical strength of the material, reduces the antigenicity, but at the same time reduces the osteoinductivity of the material; although the decalcification treatment better retains the inductive, but greatly reduces mechanical strength

Method used

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  • Construction method of sclerotomal cell idiosyncratic transcription factor - 2 gene decorated tissue engineered bone
  • Construction method of sclerotomal cell idiosyncratic transcription factor - 2 gene decorated tissue engineered bone

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Experimental program
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Effect test

Embodiment 1

[0018] 1. Preparation of seed cells:

[0019] Draw about 2ml of mouse bone marrow suspension, centrifuge through density gradient, collect the mononuclear cell layer at the interface, adjust to 2×10 5 / cm 2 density inoculation. After 24 hours, the medium was changed for the first time, and non-adherent cells were discarded, and the medium was changed twice a week thereafter. After 14 days, the monolayer cells were basically confluent. 0.25% trypsin digestion, according to 1:2 passage. The 1st to 6th passage cells are used, and the cell confluency is about 90%, and the adenovirus carrying the human Osf2 gene is used to infect, and the cells are collected 1 day after the infection for compounding with the scaffold material.

[0020] 1.1 Acquisition of bone marrow mesenchymal stem cells: extract about 2ml of bone marrow suspension from the femur of the mouse, slowly add it to a centrifuge tube pre-added with 5mL of Percoll separation solution (specific gravity 1.073g / mL), cen...

Embodiment 2

[0033] Rabbit bone marrow mesenchymal stem cells were obtained according to step 1 in Example 1, and after Osf2 gene modification, they were planted on the scaffold material prepared according to step 2, followed by dynamic culture in vitro for 5 days according to step 3. Forty Japanese white rabbits aged 3 months were selected to establish a 1.2 cm unilateral radial defect, and were divided into 4 groups according to different repair methods, with 10 rabbits in each group: Group A. Osf2 gene-modified MSCs were repaired after compounding with acellular bone matrix material; B Group. Non-gene-modified MSCs combined with acellular bone matrix materials for repair; Group C. Repair with simple scaffold materials; Group D. Blank control without any bone grafting treatment. See results figure 2 X-ray examination and histological observation showed that group A was significantly better than other groups in terms of new bone formation, bone remodeling and medullary canal recanalizati...

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Abstract

Disclosed is a construction method of sclerotomal cell idiosyncratic transcription factor - 2 gene decorated tissue engineered bone through employing a natural biologically derived material (acellular bone matrix) and seed cells (osteoblast specific transcription factor-2 gene modified mesen chymal stem cells), carrying out extracorporal dynamic culture and restoration for large segment long bone defection. The method can realize the clinic cicatrisation of bone defections.

Description

technical field [0001] The invention relates to a method for constructing tissue-engineered bone modified by osteoblast-specific transcription factor-2 gene, and belongs to the technical field of bone tissue engineering. Background technique [0002] Bone defects caused by tumor resection, trauma, bone disease and abnormal bone growth are difficult to heal only by self-repair, so bone grafting is the most effective measure for treating large bone defects. Bone grafting has become second only to blood transfusion in tissue transplantation in the United States. According to incomplete statistics from the Centers for Disease Control and Prevention in the United States, more than 1 million patients need bone graft surgery due to bone defects in the United States every year, and the medical expenses exceed 5 billion US dollars, and the cost in the United Kingdom also exceeds 900 million pounds. The traditional transplantation methods mainly include aut...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38
Inventor 董世武应大君谢肇
Owner ARMY MEDICAL UNIV
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