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Cartilage complex of tissue engineering bone and method for preparing same

A tissue engineering bone and composite technology, applied in prosthetics, medical science, etc., can solve the problems of slow degradation of the underlying hydroxyapatite, complicated operation, and ineffective fixation of tissue engineered articular cartilage products.

Inactive Publication Date: 2010-09-01
中国人民解放军第三0九医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

(2) The fixation problem of tissue engineered articular cartilage products has not been effectively solved
(3) Existing tissue engineered cartilage has biomechanical defects
(4) Poor integration between tissue engineered cartilage and subchondral bone
[0005] The composite osteochondral scaffold using the natural gel-like material as the upper cartilage scaffold has the following problems: ①The upper layer material has poor mechanical properties; ②There are mechanical defects between the interfaces, which are easy to tear off and have poor shear resistance; ③Gel-like scaffolds After implanting in the body, it absorbs water and deforms, turning into a gel again, and the mechanical properties of the cartilage layer are further weakened; ④The operation is relatively complicated; ⑤The lower layer of hydroxyapatite degrades slowly, which affects the healing with the surrounding bone
[0010] There are many studies and patents on the application of decalcified bone matrix as a scaffold for cartilage tissue engineering, but they are all only used for cartilage tissue engineering, and the aforementioned fixation problems still exist when implanted in the body

Method used

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  • Cartilage complex of tissue engineering bone and method for preparing same
  • Cartilage complex of tissue engineering bone and method for preparing same

Examples

Experimental program
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Effect test

Embodiment 1

[0044] Embodiment 1 uses fresh bovine femoral condyle as raw material to prepare tissue engineered osteochondral scaffold

[0045] Purchasing fresh bovine femoral condyles from the market, removing the attached muscles and ligaments, sawing them into 2cm thick bone slices, washing the bone marrow with a high-pressure water gun and triple distilled water, cutting off the bone cortex, and dehydrating with absolute ethanol , degreased with ether, and air-dried. Then a trephine with a diameter of 1 cm was used to make a cancellous bone column with a diameter of 1 cm from the treated bovine femoral condyle. The lower 2 / 3 of the cancellous bone column was embedded in paraffin. The lower 2 / 3 part of the paraffin-embedded cancellous bone column was placed in 10% (w / v) EDTA solution for decalcification for 2 weeks, then the paraffin was removed with xylene and graded alcohol, and soaked in PBS buffer for more than 1 week. Then freeze-dry to obtain the tissue engineering osteochondral...

Embodiment 2

[0046] Example 2 HE staining to identify the decalcification effect of tissue engineered osteochondral scaffold

[0047] The tissue-engineered osteochondral scaffolds prepared in Example 1 were respectively resin-embedded, sectioned into hard tissues, stained with HE, and placed under a macroscopic microscope for photographing and observation, as shown in figure 1 shown. The results showed that the periphery of the upper layer of the stent (i.e. the upper 1 / 3 part of the stent) and the lower layer of the stent (i.e. the lower 2 / 3 paraffin-embedded part of the stent) was stained light red, and the lower layer of the stent (i.e. the lower 2 / 3 paraffin-embedded part of the stent) The central area of ​​the embedding part) was stained purple-blue, and the interface between the upper and lower layers was very clear.

[0048] It can be seen that, through the layered decalcification treatment of the present invention, the upper layer (i.e. the upper 1 / 3 part) of the prepared tissue e...

Embodiment 3

[0049] Example 3 Sodium Alginate Gel as Secondary Scaffold to Construct Tissue Engineering Osteochondral Complex

[0050] Collect the second-generation rabbit chondrocytes cultured and expanded by conventional methods, and dilute them to 4×10 with 1.2% (v / w) sodium alginate solution. 7 / ml, get 0.5ml above-mentioned cell suspension and join in the hole of 48-well culture plate, the layered tissue engineered osteochondral scaffold prepared in embodiment 1 is inverted, allow the upper layer of the scaffold (that is, the completely decalcified part of the scaffold) and only the upper layer Immerse in the solution in the above-mentioned culture dish or culture plate, the lower layer of the support (i.e. only the decalcified part of the surface of the support) is exposed to the solution, and the liquid surface is 5mm away from the decalcification interface of the support (i.e. the interface between the upper and lower layers of the support), Add 0.1ml 2% (w / v) CaCl 2 Solution, put...

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Abstract

The invention belongs to the field of cartilage tissue repair and tissue engineering and provides a cartilage complex of a tissue engineering bone and a method for preparing the same. The preparation method comprises the following steps of: performing marrow removal, degreasing and layered calcium-removal of cancellous allograft bones or xenogeneic cancellous bones to obtain the cartilage framework of tissue engineering bones, wherein the upper layer of the cartilage framework is completely decalcified and only the surface of the lower layer is decalcified; and using the framework as a primary framework, adding gel-type matters, such as fibrinogens or alginic acid, serving as a secondary framework and a distribution carrier to obtain the cartilage complex of the tissue engineering bone. Compared with the conventional tissue engineering bone cartilage, the cartilage complex of the invention has the advantages of complete natural biomaterials, complete integration, no application of the layered stacking and interface combination technology, high pressure resistant, tensile resistant and shearing resistant properties, uniform and accurate loading of cells, effective enforcement of the differentiation of seeded cells into cartilage cells, easy operation, convenient minimally invasive operation under an arthroscope, and the like.

Description

technical field [0001] The invention belongs to the field of cartilage tissue repair and tissue engineering. The invention provides a method for constructing tissue engineering osteochondral complex. Specifically, firstly, the layered decalcification technology was used to prepare tissue-engineered osteochondral scaffolds with the cancellous bone column as the main body, the upper layer was completely decalcified, and the lower layer was only superficially decalcified. , using fibrinogen gel or alginate gel as the secondary scaffold and distribution carrier of cartilage seed cells to construct tissue engineered osteochondral complex. Background technique [0002] Injuries and lesions of articular cartilage are common diseases, and it is difficult to repair by itself. Tissue-engineered cartilage is the most promising repair method. However, the following problems need to be solved urgently in the research of tissue engineering of articular cartilage: (1) When repairing art...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/36A61L27/26A61L27/22A61L27/20A61L27/52
Inventor 余方圆黄利虹马远征刘海容
Owner 中国人民解放军第三0九医院
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