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A magnetically responsive tissue engineering material capable of promoting osteogenesis, its preparation method and application

A technology of tissue engineering and osteogenesis, applied in tissue regeneration, prosthesis, medical science, etc., can solve the problems of regulating and regulating the ability of macrophages to polarize to M2, the disorder of fiber structure arrangement, etc., to increase osteogenesis effect of effect

Active Publication Date: 2022-03-11
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] In order to overcome the shortcomings of existing tissue engineering materials (such as hydrogel, etc.) that the fiber structure is disorderly arranged, it is impossible to regulate in real time according to the experimental requirements, and the ability to regulate the polarization of macrophages to M2 is weak, etc., the present invention provides a new type of "geni Flat cross-linked Fe3O4 magnetic particle collagen I hydrogel" magnetic response tissue engineering material

Method used

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  • A magnetically responsive tissue engineering material capable of promoting osteogenesis, its preparation method and application
  • A magnetically responsive tissue engineering material capable of promoting osteogenesis, its preparation method and application
  • A magnetically responsive tissue engineering material capable of promoting osteogenesis, its preparation method and application

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preparation example Construction

[0030] Such as figure 1 It is a schematic flow chart of the preparation method of the magnetic response tissue engineering material with osteogenesis-promoting effect of the present invention, and the specific operation steps are as follows:

[0031] Step 1 Prepare reagents: add 1ml 1× phosphate buffered saline (PBS) to 0.01g of genipin to make 1% genipin solution, put it in a constant temperature incubator at 37°C for 3 days in the dark, and dissolve it completely After that, use a bacteria filter to filter the bacteria, and store them in a refrigerator at 4°C in the dark; 10xPBS buffer solution, 1mol / L sodium hydroxide (NaOH) solution, 0.1mol / L NaOH solution and 0.1mol / L hydrochloric acid (HCl) solution are all filtered After filtering the bacteria, put it in a 4°C refrigerator for later use; the collagen I solution was directly placed in a 4°C refrigerator for later use. Special cell culture plates, EP tubes, tweezers and tissue scissors are sterilized by high-pressure ste...

Embodiment 1

[0042] 2D culture of macrophages: soak cell slides (φ14mm) in 95% ethanol solution for 30 minutes to sterilize, pick them up with tweezers and let them dry naturally in an ultra-clean bench, then put them into a special cell culture plate; Use tissue scissors to cut off a small section of the pipette tip with a size of 1ml or 200μ (depending on the actual amount of glue added), and take the genipin cross-linked ferroferric oxide magnetic About 100-300 μL of granular collagen I hydrogel is placed on the cell slide in a special cell culture plate, and incubated in a 37°C cell culture incubator for 3 hours until it forms a gel on the surface of the slide; then according to the actual experimental needs, the material in each well Add 100,000-1,000,000 primary macrophages on the surface, and add an appropriate amount of macrophage primary culture medium, place Figure 4 The in vitro culture magnetic field mold shown above can be cultured in a 37°C constant temperature incubator for...

Embodiment 2

[0044] 3D culture of macrophages: Soak cell slides (φ14mm) in 95% ethanol solution, pick them up with tweezers and let them dry naturally in an ultra-clean bench, then put them into special cell culture plates for later use. Take the unincubated genipin cross-linked ferroferric oxide magnetic particle collagen I pregel solution described in step 5 and place it in a 37°C cell culture incubator for 30 minutes. At this time, the material properties are slightly light blue gel-like (already Initially gelled, but not yet shaped). According to the ratio of 100,000-1 million primary macrophages per 100-300 μL of material, add the primary macrophage suspension to the EP tube containing the material at a volume ratio of 1:1; Put a small section of the pipette tip into the EP tube with the trimmed pipette head and gently blow it several times to make the cell suspension and pre-gel solution fully mixed, and draw about 100-300μL with the same clean pipette head Put the material on the c...

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Abstract

The invention discloses a magnetic responsive tissue engineering material with promoting osteogenesis and a preparation method and application thereof. The preparation method of the material is as follows: firstly adjusting the pH value and osmotic pressure of a collagen I solution, and then adding sequentially in a certain proportion The aminated ferroferric oxide magnetic particle solution and the cross-linking agent genipin solution need to be shaken and mixed evenly after each addition of the solution, and finally incubated in a constant temperature incubator at 37°C for at least 3 hours to obtain genipin cross-linking Ferric oxide magnetic particle collagen I hydrogel. The material can change the morphology of its internal fibrous structure temporally under the action of an external magnetic field, so that it has the ability to directionally induce the polarization of macrophages to M2, promote the osteogenesis of bone defect sites, and can be used for tissue research. Effects of engineered materials on the osteogenic immune microenvironment and its mechanism.

Description

technical field [0001] The invention belongs to the technical field of biomedical materials. More specifically, it relates to a magnetically responsive tissue engineering material capable of promoting osteogenesis, its preparation method and application. Background technique [0002] Large bone defects are often caused by trauma, bone tumor resection, and severe infection. The treatment cycle is long, difficult, with many complications and high disability rate. It is a major technical difficulty to be solved in orthopedics clinical practice. The current common treatment methods include autologous bone graft, allogeneic bone graft and bone tissue engineering. Among them, the amount of autologous bone is limited, which cannot meet the requirements of large defect bone transplantation, and will cause new trauma to the bone donor area. However, allograft bone grafts have problems such as graft rejection, easy transmission of diseases, and high rate of bone nonunion. Therefore...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/24A61L27/02A61L27/52A61L27/50
CPCA61L27/24A61L27/025A61L27/52A61L27/50A61L2430/02C08L89/00
Inventor 叶招明黄东骅徐恺成黄鑫林秾俞小华
Owner ZHEJIANG UNIV
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