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Production method of finished-product tissue engineered bone

A technology of tissue engineering and finished products, applied in medical science, prosthesis, etc., can solve the problems that the sterility of tissue-engineered bone cannot be well guaranteed, the construction period takes a long time, and rejection reactions, etc., to facilitate long-term storage , high integrity, and improved safety

Inactive Publication Date: 2015-09-02
SHANDONG UNIV QILU HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the main reasons hindering the clinical application of tissue-engineered bone constructed in vitro include: (1) the construction period is relatively long (about 1 month), and the process is relatively complicated. , will not wait for the construction of tissue-engineered bone after receiving patients, and patients are also unwilling to wait; (2) The constructed tissue-engineered bone has certain immunogenicity, and there may be rejection reactions when implanted in the human body; ( 3) The sterility of tissue engineered bone constructed in the laboratory cannot be well guaranteed

Method used

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  • Production method of finished-product tissue engineered bone

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Such as figure 1 as shown,

[0034] (1) Firstly, the bone marrow was extracted from the iliac crest of volunteers, and bone marrow mononuclear cells were obtained by adherent culture method, and expanded and cultured. The flow cytometry method detects the marker antigen on the cell surface, which can confirm that the obtained cells are bone marrow mesenchymal stem cells. The third to sixth generation cells were applied to the construction of tissue engineered bone in vitro.

[0035] (2) The cells obtained by subculture were made into a single cell suspension, and the cell concentration was controlled at 2.0×10 6 / ml or so. Then infiltrate the porous β-TCP scaffold into the single cell suspension, and use the method of vacuuming (vacuum degree is 25.33Kpa) to make the single cell suspension fully enter the interior of the porous scaffold. Cultivate in the cell incubator for 2 hours to obtain the cell / scaffold complex, then inoculate the cell / scaffold complex into the...

Embodiment 2

[0045] Such as figure 1 as shown,

[0046] (1) Firstly, the bone marrow was extracted from the ilium of volunteers, and the bone marrow mononuclear cells were obtained by density gradient centrifugation, and then expanded and cultured. The third to sixth generation cells were applied to the construction of tissue engineered bone in vitro.

[0047] (2) The cells obtained by subculture were made into a single cell suspension, and the cell concentration was controlled at 2.0×10 7 / ml. Then the porous hydroxyapatite scaffold is soaked into the single cell suspension, and the single cell suspension is fully entered into the interior of the porous scaffold by vacuuming, and the vacuum degree is 75.99Kpa. Then cultivate in the cell culture incubator for 4 hours to obtain the cell / scaffold complex, inoculate the cell / scaffold complex into the perfusion bioreactor, and add the medium After standing for 4 hours, start the perfusion bioreactor , the cell / scaffold complex was continuou...

Embodiment 3

[0057] Such as figure 1 as shown,

[0058] (1) Firstly, the bone marrow was extracted from the ilium of volunteers, and the bone marrow mononuclear cells were obtained by density gradient centrifugation, and then expanded and cultured. The third to sixth generation cells were applied to the construction of tissue engineered bone in vitro.

[0059] (2) The cells obtained by subculture were made into a single cell suspension, and the cell concentration was controlled at 2.0×10 7 / ml. Then soak the porous coral support into the single-cell suspension, and use a vacuum method to make the single-cell suspension fully enter the inside of the porous support, and the vacuum degree is 55Kpa. Then cultivate in the cell culture box for 3 hours to obtain the cell / scaffold complex, inoculate the cell / scaffold complex into the perfusion bioreactor, and add the culture medium to start the perfusion bioreactor after standing for 3 hours, The cell / scaffold complex was continuously perfused...

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Abstract

The invention relates to a production method of a fished-product tissue engineered bone. The method includes: acquiring the mononuclear cell of marrow, and performing multiplication culture to obtain single-cell suspension with the concentration of 2.0*10<6>-2.0*10<7> / ml; soaking a porous scaffold into the single-cell suspension to allow the single-cell suspension to enter the porous scaffold sufficiently, and culturing to obtain cell / scaffold composite; inoculating the cell / scaffold composite into a perfusion type bioreactor, adding culture medium and standing for 2-4 hours, and then performing continuous perfusion culture on the cell / scaffold composite to obtain the tissue engineered bone; performing preliminary cleaning, cryopreservation, virus inactivation, ultrasonic cleaning, vacuum freezing drying, aseptic packaging and radiation sterilizing on the tissue engineered bone to obtain the finished-product tissue engineered bone. The method has the advantages that the cryopreservation is performed on the produced tissue engineered bone, and the immunogenicity of the tissue engineered bone can be effectively lowered.

Description

technical field [0001] The invention specifically relates to a preparation method of a finished tissue-engineered bone. Background technique [0002] Due to various reasons, especially the repair and reconstruction of large bone defects caused by severe trauma, large bone tumor resection, and joint prosthesis loosening, it is a major problem in orthopedic treatment. At present, the methods for treating bone defects mainly include autologous bone grafting, allogeneic bone grafting, artificial bone materials, and metal implants. However, autologous bone transplantation will have complications such as limited bone volume, infection at the donor site, and chronic pain; allogeneic bone transplantation includes allogeneic bone grafts and xenogeneic bone grafts, but there are immune rejection, insufficient strength, and disease transmission. Complications. With the improvement of laws, donors must be donated by patients, and their sources are becoming less and less; while artifici...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/12A61L27/56A61L27/54
Inventor 李德强李明刘培来张元凯李建民
Owner SHANDONG UNIV QILU HOSPITAL
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