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Ferroferric oxide superparamagnetic nanoparticle stimulating stem cell for exosome osteogenesis

A superparamagnetic and nanoparticle technology, which is applied in the field of nanomaterials and biomedicine, can solve the problems of seed cell apoptosis, poor osteogenesis effect, poor stability, etc.

Active Publication Date: 2019-05-14
PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the lack of blood supply in the host microenvironment, seed cells are prone to apoptosis in a hypoxic environment, resulting in poor osteogenesis
However, it has problems such as low survival rate, poor stability, immune rejection, tumorization in vitro stimulation culture, etc., which seriously limit its wide clinical application.

Method used

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  • Ferroferric oxide superparamagnetic nanoparticle stimulating stem cell for exosome osteogenesis
  • Ferroferric oxide superparamagnetic nanoparticle stimulating stem cell for exosome osteogenesis
  • Ferroferric oxide superparamagnetic nanoparticle stimulating stem cell for exosome osteogenesis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1 The cultivation of bone marrow mesenchymal stem cells (BMSCs)

[0042] The human bone marrow mesenchymal stem cells used in the experiments of the present invention were purchased from Shanghai Youyu Biotechnology Co., Ltd.

[0043] The BMSCs cells were subcultured with low-sugar DMEM medium containing 10% fetal bovine serum at a ratio of 1:3 to obtain the first generation of bone marrow mesenchymal stem cells, and continued to be subcultured to the third generation of bone marrow mesenchymal stem cells. In this experiment, the cells of passage 3-5 were selected for subsequent experiments.

Embodiment 2

[0044] Example 2 CCK-8 detects Fe 3 o 4 Toxic to bone marrow mesenchymal stem cells

[0045] The 3rd-5th generation BMSCs were inoculated in 96-well plates, and the planting density of the cells was 1×10 4 / well, after 2 days of culture, replace with Fe 3 o 4 culture medium, adding concentrations of 0mg / ml, 0.5mg / ml, 1mg / ml, 2mg / ml Fe 3 o 4 , treatment 5d, using the CCK8 method for Fe 3 o 4 Cells treated for 1d, 2d, 3d and 5d were tested for toxicity. Add 10 μL of CCK8 solution and 90 μL of DMEM medium to each well, and measure the absorbance of each well with a microplate reader, and the absorption wavelength of the CCK8 solution is 450 nm.

[0046] CCK8 cytotoxicity experiments found that using a concentration gradient of 0-1mg / ml Fe 3 o 4 There is no obvious toxic effect on BMSCs, Fe at a concentration greater than 1mg / ml 3 o 4 have a certain effect on cell proliferation ( figure 1 ).

Embodiment 3

[0047] Example 3 Fe 3 o 4 Superparamagnetic nanoparticles stimulate exosomes from bone marrow mesenchymal stem cells

[0048] Take the well-grown BMSCs of the 3rd-5th generation and press 1×10 5 Seed in a 6-well plate at a density of 6 wells. When the cell fusion rate reaches 70-80%, divide the cells into 2 groups: control group and Fe 3 o 4 treatment group. Control group: cultivated with 10% fetal bovine serum low-sugar DMEM medium; Fe 3 o 4 Treatment group: Fe 3 o 4 Superparamagnetic nanoparticles (Nanjing Xiaoli Biology) were dissolved at 1 mg / ml in low-sugar DMEM culture medium containing 10% fetal bovine serum by volume, and the cells were cultured.

[0049] 1. Observation of Fe by transmission electron microscope 3 o 4 Mark BMSCs cells:

[0050] Fe 3 o 4 Treated BMSCs at 37°C, 5% CO 2 Cultured in an incubator for 48 hours, digested with 0.25% trypsin at 37°C for 1 minute, centrifuged at 120 g for 5 minutes, washed twice with PBS, fixed with 2.5% glutaraldeh...

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Abstract

The invention discloses an exosome preparation derived from a stem cell. The exosome preparation is an exosome secreted by a Fe3O4 superparamagnetic nanoparticle stimulating the stem cell. The invention further provides the application of the exosome preparation in preparing a pharmaceutic preparation for promoting mesenchymal stem cell osteogenic differentiation and the application of the exosomepreparation in preparing a biological agent for treating orthopedic disorders or bone repair materials. In consideration of the complicated pathophysiologic process of osteonecrosis and bone defects,the exosome preparation prepared by the exosome secreted by the Fe3O4 superparamagnetic nanoparticle stimulating the stem cell is adopted, intramuscular administration can be adopted at local bone defect parts, or local filling is performed after the exosome preparation is mixed with absorbable gel, the exosome preparation directly reaches the focus, the osteogenic activity of the mesenchymal stem cell is motivated, the deposition of osteoblasts is promoted, and the repair of the osteonecrosis or bone defects is promoted.

Description

technical field [0001] The invention relates to the field of nanomaterials and biomedical technology, in particular to Fe 3 o 4 Superparamagnetic nanoparticles stimulate bone marrow mesenchymal stem cell exosome osteogenesis. Background technique [0002] Bone tissue defect is caused by factors such as trauma, infection, tumor, and abnormal bone development, which cause the body to lose some bone mass and form a large gap. This problem is relatively common in clinical practice and difficult to deal with. How to promote the repair of bone tissue defects has always been the focus of research by researchers in this field. Although autologous bone transplantation is the best method of treatment, autologous bone source transplantation is extremely limited and increases the suffering of patients, while allogeneic bone transplantation has immune reactions and many complications. [0003] In recent years, with the development of bone tissue engineering technology, this problem ha...

Claims

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Application Information

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IPC IPC(8): A61K35/28A61L27/54A61L27/36A61L27/52A61P19/08C12N5/0775
Inventor 王海于凌佳吴狄贺宇吴志宏
Owner PEKING UNION MEDICAL COLLEGE HOSPITAL CHINESE ACAD OF MEDICAL SCI
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