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Injectable tissue-engineered bone repair material and construction method thereof

A tissue engineering bone and repair material technology, applied in the field of biomedical engineering, can solve the problems of affecting the osteogenic activity of seed cells, insufficient oxygen supply, etc., to overcome poor hypoxia resistance, improve osteogenic ability, and overcome low seeding density Effect

Inactive Publication Date: 2019-06-21
HANGZHOU DANBINNI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When the seed cells are aggregated together in a certain form, due to the tight connection between the cells, the seeding density can be significantly increased and can reach 10 8 However, in the construction of tissue engineering bone repair materials, the seeding density of seed cells is much higher than that of conventional cell culture, so seed cells are prone to insufficient oxygen supply in tissue engineering bone repair materials, thus Severely affects the osteogenic activity of seeded cells

Method used

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  • Injectable tissue-engineered bone repair material and construction method thereof
  • Injectable tissue-engineered bone repair material and construction method thereof
  • Injectable tissue-engineered bone repair material and construction method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1, the cultivation of bone marrow stromal stem cells and the construction of cell microspheres

[0039] The culture of rat bone marrow stromal stem cells: 6-week-old healthy SD rats were killed, and the bones of both lower limbs were taken out, and the bone marrow was washed out under sterile conditions. 2 Cultivate in an incubator, change the medium every three days, and digest and passage after the primary cells grow to the bottom of the bottle.

[0040] Construction of cell microspheres: cell microspheres were obtained by rotating culture method, and the cell density was (2-5)×10 5 The suspension of bone marrow stromal stem cells per ml was placed in a rotary culture flask, and placed on a magnetic stirrer for 16-24 hours to rotate and cultivate at a speed of 25-80 rpm to obtain a cell density of 10 8 Order of magnitude bone marrow stromal cell microspheres. However, it is not limited thereto, and it can also be cultured by the hanging drop method.

[0041] ...

Embodiment 2

[0046] Embodiment two, the construction of injectable gel carrier

[0047] Sodium alginate and fibrinogen are mixed at a volume ratio of 1:3 to form a gel carrier, wherein the mass concentration of sodium alginate is 0.1%-4%, and the concentration of fibrinogen is 5-30mg / ml.

[0048] Preferably, sodium alginate and fibrinogen are sterilized by gamma rays to prepare sodium alginate with a mass concentration of 1% and fibrinogen with a concentration of 20 mg / ml, and then 1% sodium alginate and 20 mg / ml ml fibrinogen was mixed at a volume ratio of 1:3 to form a gel carrier.

Embodiment 3

[0049] Example three, the survival rate of single cells and cell microspheres of different cell densities in the gel carrier

[0050] The cell density was 0.5 × 10 7 / ml, 1×10 7 / ml, 2×10 7 / ml, 5×10 7 / ml, 10×10 7 / ml and 20×10 7 / ml of single cells and cell microspheres were inoculated in the gel carrier, the volume of the gel carrier is 0.5ml, placed in a 6-well cell culture plate, added 10ml of DMEM culture solution containing 10% fetal bovine serum, every 3 The culture medium was replaced every day, and cultured for 14 days. After 14 days, the gel was broken and the cells were collected, and the cell viability was detected by the trypan blue exclusion method. For test results, see Figure 5 , Figure 5 It is a comparison chart of the survival rate of single cells and cell microspheres in the gel carrier. Under the same cell density, when the cell density exceeds 1×10 7 / ml, the survival rate of the cell microspheres in the gel carrier far exceeds that of a single ...

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Abstract

The application discloses an injectable tissue-engineered bone repair material and a construction method thereof, wherein the injectable tissue-engineered bone repair material comprises a gel carrierand seed cells; the seed cells are attached to the gel carrier to form a complex with bone tissue three-dimensional structure and physiological activity; the gel carrier is made from injectable sodiumalginate and fibrinogen mixture and used for combining with the seed cells; the seed cells are bone marrow stroma stem cell microspheres. Single cells are replaced with the cell microspheres to act as the seed cells, so that the problem that single cells have low inoculation density and poor anoxia resistance is solved; therefore, osteogenic capability of the injectable tissue-engineered bone repair material is improved.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to an injectable tissue engineering bone repair material and a construction method thereof. Background technique [0002] Injectable tissue engineering bone repair materials are usually compounded with hydrogel-like scaffold carriers, seed cells, and active factors in a certain way, injected into the body, and transformed into a solid state through physical or chemical action in the body. Or fixed in some way, the implant gradually generates bone tissue and plays a role in bone repair. [0003] Among the components of injectable tissue engineered bone repair materials, seed cells play a central role in bone formation. The number, density and osteogenic activity of seed cells determine the osteogenesis ability of injectable tissue engineering bone repair materials. The existing common seed cells exist in the form of single cells in the injectable tissue engineering b...

Claims

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Application Information

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IPC IPC(8): A61L27/52A61L27/20A61L27/22A61L27/38
Inventor 刘玉侃蔡玉荣
Owner HANGZHOU DANBINNI BIOTECH
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