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Construction method of tissue engineering bone repair material

A tissue engineering bone and repair material technology, applied in the direction of bone/connective tissue cells, medical science, prosthesis, etc., can solve the problem of lack of biomechanical strength, and achieve good biodegradability, low cost, and complete degradation and absorption. Effect

Inactive Publication Date: 2013-06-19
XIAN REJE BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the materials used for injectable bone are gels, mainly including fibrin gels, polyethylene oxide hydrogels, alginate gels, etc. These biomaterials are gel-like after being injected into the body and do not have Biomechanical strength, although the composite seed cells have a certain osteogenic effect after implantation in the body, there is still a considerable distance from the ideal injectable bone scaffold material

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Including the following steps:

[0021] 1) Cultivation of seed cells: Bone marrow was extracted, and bone marrow stromal stem cells were isolated and cultured by the whole bone marrow culture method. The medium used was DMEM medium containing 10% fetal bovine serum. The cells were subcultured and proliferated until the third generation 10 7 Magnitude;

[0022] 2) Preparation of injectable scaffold material: compound chitosan and β-tricalcium phosphate at a ratio of 1ml:0.5g in vitro to form a chitosan-β-tricalcium phosphate complex;

[0023] 3) Treatment of seed cells: culture to the third generation 10 7 Bone marrow stromal stem cells of order of magnitude were mixed with 0.3ml of DMEM;

[0024] 4) Compounding of seed cells and chitosan-β-tricalcium phosphate: After 8 minutes of compounding of chitosan-β-tricalcium phosphate, immediately add the seed cells treated in step (3), the addition ratio is chitosan, 3ml: 1.5g: 0.3ml of β-tricalcium phosphate and DMEM are th...

Embodiment 2

[0027] After inoculating the cells, discard the original medium on the 2nd, 4th, 6th, and 8th day, add 1ml of serum-free DMEM and 10μl of MTT (5mg / mol), and incubate at 37°C in CO 2 Incubate in the incubator for 4 hours, carefully absorb MTT, add 0.5ml of dimethyl sulfoxide and shake for 15 minutes, select a wavelength of 570nm on the microplate reader, and measure the light absorption value.

[0028] With the prolongation of culture time, the D570 of each group gradually increased, reaching the highest level on the 8th day, and there was no significant difference between the chitosan-β-tricalcium phosphate group and the control group at each time point (>0.05) (Table 1).

[0029] Table 1 Absorbance values ​​measured by two groups of MTT method

[0030] Group 2 days 4 days 6 days 8 days control group 0.120±0.027 0.404±0.041 0.902±0.097 1.197±0.074 Chitosan-β-tricalcium phosphate 0.116±0.021 0.372± 0.05 0.87±0.06 1.139±0.094

[0031] P>0...

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Abstract

The invention discloses a construction method of a tissue engineering bone repair material. The method includes: 1) culture of seed cells; 2) preparation of an injectable scaffold material; 3) treatment of the seed cells; and 4) compounding of the seed cells and chitosan-beta-tricalcium phosphate, and other steps. The bone repair material provided in the invention is composed by compounding an injectable material with bone marrow stroma stem cells, can be arbitrarily shaped and cell compounding is simple. Being able to be completely degraded and absorbed within certain period of time, the material has an obvious inductive osteogenesis effect, and the cost is low.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering, and in particular relates to a construction method of a tissue engineering bone repair material. Background technique [0002] Tissue engineering is a hot spot in life science in the 21st century. Its core content is three-dimensional cell culture in vitro, and its basic method is to form a complex of seed cells and biodegradable biomaterials to replace damaged tissues and organs. Injectable tissue-engineered bone is a liquid-state injectable tissue-engineered bone that is prepared by using degradable biomaterials that are initially liquid and organically compounded with seed cells, and gradually solidifies to form a gel or gel during the process of completely filling the bone defect. Solid-state tissue-engineered bone for bone defect repair. Injectable bone is easy to shape, easy to compound with seed cells and growth factors, and has the advantages of less tissue damage, no damage ...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/20C12N5/0775
Inventor 孙叶芳
Owner XIAN REJE BIOLOGICAL TECH
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