A kind of preparation method of magnetic control protein composite cell membrane sheet

A cell membrane, protein technology, applied in pharmaceutical formulations, prostheses, drug delivery, etc., can solve problems that have not been reported in literature, and achieve the effect of promoting bone regeneration in vivo, high saturation magnetization, and improving the effect of regeneration and repair in vivo.

Active Publication Date: 2020-09-01
SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to further improve the effect of regeneration and repair, it is planned to incorporate growth factors with different biological activities into the cell membrane to construct a magnetically controlled growth factor composite cell membrane. This aspect of research has not yet been reported in the literature.

Method used

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  • A kind of preparation method of magnetic control protein composite cell membrane sheet
  • A kind of preparation method of magnetic control protein composite cell membrane sheet
  • A kind of preparation method of magnetic control protein composite cell membrane sheet

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1. nGO@Fe 3 o 4 Preparation and Characterization of Magnetic Nanoparticles

[0029] Step 1. Preparation of nano-sized graphene oxide (nGO)

[0030] according to figure 1 As shown in a, in order to prepare nGO@Fe3O 4 For magnetic nanoparticles, nanometer-sized graphene oxide is firstly prepared. 20mL of AgNO 3 The solution was added to an equal volume of 0.5mg / mL graphene oxide solution, stirred at room temperature for 30 minutes, and then left to stand for 48 hours (AFM characterization results are as follows: figure 1 As shown in b, graphene oxide particles vary in size). Centrifuge at 2000 rpm for 10 minutes, collect the supernatant and mix it with 10 mL of nitric acid solution (2 mM), concentrate in a rotary vacuum evaporator for 5 hours to dissolve the silver particles. The concentrated solution was centrifuged at 6000rpm for 10 minutes to remove the precipitate containing large pieces of graphene oxide. The supernatant was further centrifuged at 100...

Embodiment 2

[0035] Example 2. Phagocytosis of nGO@Fe by stem cells 3 o 4 Magnetic nanoparticles for cell labeling

[0036] Step 1. Cells and nGO@Fe 3 o 4 Magnetic nanoparticle co-culture

[0037] Use serum-free DMEM culture medium to prepare a 200 μg / mL magnetic nanoparticle solution for later use. Dental pulp stem cells were extracted from clinically impacted teeth. When the dental pulp stem cells were cultured to about 50% confluence, the serum-free DMEM medium was replaced, and magnetic nanoparticles with final concentrations of 0, 1, 5, 10, 50, and 100 μg / mL were added to co-culture for 2 hours, and observed by Prussian blue staining Phagocytosis of granules by cells. Such as image 3 As shown in a, with the increase of particle concentration, cells phagocytized more particles, which were mainly distributed around the nucleus.

[0038] Step 2. Detection of cell apoptosis and cell proliferation activity

[0039] When the dental pulp stem cells were cultured to about 80% conflu...

Embodiment 3

[0040] Example 3. nGO@Fe 3 o 4 Magnetic control effect of magnetic nanoparticles labeling stem cells

[0041] Step 1. Cells and nGO@Fe 3 o 4 Magnetic nanoparticle co-culture

[0042] The procedure for cell labeling was the same as in Example 2: when the dental pulp stem cells were cultured to about 80% confluency, the serum-free DMEM medium was replaced, and magnetic nanoparticles with a final concentration of 100 μg / mL were added for co-cultivation for 2 hours. Such as Figure 4 As shown in a, the cells labeled with magnetic particles were added to the culture medium, and adsorbed and immobilized under the control of a magnet for 4 hours to form a cell membrane.

[0043] In addition to dental pulp stem cells, we also used other cells that can also be labeled with magnetic particles, such as the NIH3T3 cell line ( Figure 5 a). At the same time, NIH3T3 cells labeled with magnetic particles can also be magnetically adsorbed to form cell membranes of specific shapes ( F...

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Abstract

The invention discloses a preparation method of a magnetic-control protein-composited cell membrane and an application of magnetic nanoparticles on preparation of the protein-composited cell membranes. The magnetic nanoparticles are dispersed with the diameter of each being 5-500nm, nanopores, chemical bonds or electric charge are contained on the surface of the magnetic nanoparticles so as to have protein attached, and the protein-composited cell membrane is established by controlling deposition of protein and cells through a magnetic field. The concept of the magnetic-control protein-composited cell membrane and the preparation method thereof are put forward, the protein can be effectively and fixedly attached to the cell membrane to play a biological role, regeneration repair effect invivo can be improved remarkably, and the novel method is provided for in vitro construction of a complex structure; the preparation for the nGO@Fe3O4 magnetic nanoparticles which are controllable anddispersing and 8-12nm in size is provided, the magnetic nanoparticles can be devoured by the cells to have the cells labeled and can adsorb the binding protein.

Description

technical field [0001] The invention belongs to the field of tissue engineering and regenerative medicine, and more specifically relates to a preparation method of a magnetically controlled protein composite cell membrane, which can be used for tissue regeneration and repair, and in vitro microtissue construction and research. Background technique [0002] The preparation of cell membranes does not require scaffold materials, and forms a whole through cell connections and extracellular matrix, which is closer to the natural physiological state of tissue cells. This technology has many advantages in stem cell-based regenerative therapy. In terms of preparation methods, the traditional use of temperature-sensitive culture dishes and direct induction film formation technology has disadvantages such as long culture period, limited cell membrane thickness, and difficulty in direct stacking of various cells. In addition, temperature-sensitive culture dishes are more expensive , wh...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/38A61L27/22A61L27/54A61L27/08A61L27/02A61L27/50
CPCA61L27/025A61L27/08A61L27/227A61L27/3804A61L27/3808A61L27/3817A61L27/3834A61L27/50A61L27/54A61L2300/412A61L2400/12C08L89/00
Inventor 蒋欣泉张文杰杨光正王贤松
Owner SHANGHAI NINTH PEOPLES HOSPITAL SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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